Source: DeepTech

Introduction

Organoids are self-assembled three-dimensional cells derived from stem cells in vitro. Although organoids are not human organs in the true sense, organoids have cell types and complex spatial morphology corresponding to stem cells in tissues and organs, and can show the interaction between cells and the surrounding matrix as well as the spatial position morphology, and they can simulate some functions and physiological reactions of tissues and organs. High similarity to the source organization.

Since the establishment of the first organoid in 2009, the field of organoids has shown a rapid development trend, and this breakthrough achievement has also been widely recognized by the scientific research community and the industry. In 2013, the journal Science named organoids among the top 10 scientific breakthroughs of the year; In 2015, MIT Technology Review listed organoid technology as one of the world's top 10 breakthrough technologies; In 2017, Nature Methods magazine named organoids Technology of the Year; In 2019, The NewEngland Journal of Medicine called it an excellent preclinical disease model.

As a basic research tool of life sciences, organoid technology has had a subversive impact on biomedicine, improving the means and methods for exploring life phenomena and medical problems in vitro, and showing great application potential in many fields, including developmental biology and disease modeling, drug screening and research and development, precision medicine, regenerative medicine, etc.

Organoid technology is of great value in basic research and has broad application prospects in the pharmaceutical industry. Based on this, the DeepTech research team conducted research on organoid technology and industry and wrote this report. This report systematically combs the development context of organoid technology, discusses the application scenarios of organoid technology, clearly understands the status quo of organoid industry, and insights into the challenges and development direction of organoid industry in the future.

01

Driven by technology, talent, policy and capital, the development pattern of the whole industry chain of organoids has initially taken shape. Organoids: simulate tissues and organs, and build "surrogates" of tissues and organs.

Organoids are self-assembled three-dimensional cells derived from stem cells in vitro. Although organoids are not human organs in the true sense, organoids have cell types and complex spatial morphology corresponding to stem cells in tissues and organs, and can show the interaction between cells and the surrounding matrix as well as the spatial position morphology, and they can simulate some functions and physiological reactions of tissues and organs. High similarity to the source organization.

Organoids should have at least three of the following characteristics:

The concept of organ-on-a-chip, which is easily confused with organoids, refers to a physiological microsystem of human organs built on a microfluidic chip, which can simulate and construct a microenvironment of tissues and organs containing complex factors such as a variety of living cells, functional tissue interface, biological fluid and mechanical force stimulation in vitro, reflecting the main structure and functional signs of human tissues and organs.

Historical evolution: organoid research has entered the fast lane

Organoid technology has successfully constructed almost all tissues and organs, and related research has entered the fast lane. As early as the 1980s, the term "organoid" was proposed, but it was not until 2009 that Hans Clevers' team cultured the first micro-intestinal organoids with small intestinal crypts and villi structures from adult stem cells derived from the mouse gut, which made the research of organoids enter the fast lane. After the rapid development of organoid technology in recent years, at present, almost all tissues and organs can construct 3D cultured organoids in vitro, including intestinal, retinal, brain, liver, kidney, stomach, lung, pancreas, colon, heart and blood vessel organoids from human pluripotent stem cells, and stomach, liver, pancreas, placenta and lung organoids from human adult tissues.

Scientific research achievements are improving year by year, and China's scientific research force ranks second in the world

The scientific research achievements in the field of organoids show a rapid growth trend. Web of Scienc e database searched for "organoid", from the successful construction of small intestine organoids by Hans Clevers, 23 papers were published in 2009, 1682 papers were published in 2022, and 7019 papers have been published so far. The scientific literature related to organoid technology has been increasing year by year. From the perspective of countries/regions that published literatures, China published 1,095 literatures, ranking second, becoming an important scientific research force in the field of organoids. The accumulation of scientific research in China will accelerate the industrialization of organoids.

Atlas of researchers in the field of organoids

Policies open the way for organoid technology

In addition to the development of organoid technology driven by scientific research accumulation and technological innovation, the introduction of relevant supportive policies has also played a role in opening mountains and expanding roads. International and domestic policies have been introduced to loosen the wide application of organoids, and the field of organoids has shown a strong momentum of development.

·America

In 2011, the National Institutes of Health (NIH), the Food and Drug Administration (FDA), and the Defense Advanced Research Projects Agency (DARPA) teamed up to create the Microphysiological Systems program, To improve the process of predicting whether a drug will be safe in humans. The plan raises organoids and organ-on-a-chip related technologies to the national strategic level for the first time. In 2022, the United States passed the FDA Modernization Act 2.0, the theme of which is to promote the reduction of the application of preclinical testing on animals and replace it with more modern scientific methods, such as organ-on-a-chip and microphysiological systems. In 2022, the US FDA approved the world's first new drug (NCT04658472) to enter clinical trials based solely on the pre-clinical data obtained from the "organoid chip" study.

·European Union

In 2020, the organoid research project entered the EU Horizon 2020 Strategic Plan. The project combines single-cell characterization and organoid techniques to validate organoids as reliable models of human biology. The project is one of six pilot initiatives funded by the EU's Horizon 2020 Framework programme to develop an open-access single cell Atlas of organoids, which will help the EU complete the foundation of the Human Cell Atlas project.

·China

In January 2021, the Ministry of Science and Technology issued the "Notice on the" 14th Five-Year "National Key Research and Development Plan of six key special projects for 2021 annual project declaration Guidelines", which listed the "organoid based malignant tumor disease model" as the first batch of key special tasks in the "14th Five-Year" national key research and development plan. The "14th Five-Year Plan" National key research and development Plan emphasizes that organoids, as a major technological breakthrough, are used in the establishment of disease models, and can be used to study the variation, heterogeneity and occurrence mechanism of stem cells in pathological states, explore new targets for disease diagnosis and treatment, and explore new strategies for diagnosis and treatment. In November 2021, the Drug Evaluation Center of the State Drug Administration issued the "Guiding Principles for Non-clinical Research and Evaluation of Gene Therapy Products (Trial)" and the "Guiding Principles for Non-clinical Research of Gene Modified cell Therapy Products (Trial)", and organoids were included in the guiding principles for the first time.

Capital elements help the development of the industry

The hot organoid technology attracts capital elements to enter the field and help the development of the industry. The establishment time of domestic organoid companies is concentrated from 2015 to 2021, and the entire industry is in the early infancy stage. Matching is that in the capital market, the financing of organoid companies is also at an early stage, most of which are in the angel round to the A round. Ketu Medical, Big Oak Technology and Chuangxin International, which were commercialized earlier, have entered the B round and Pre-B round after several rounds of financing. From the perspective of the primary market, in 2022, when the capital market meets the winter, the financing situation of the organoid industry is more optimistic, and the rounds are from angel to round B, with the amount ranging from tens of millions to hundreds of millions of yuan (incomplete statistics).

A development pattern of the organoid industry chain has taken shape

At present, the development pattern of the whole domestic organoid industry chain has taken shape, and the upper, middle and lower reaches have been distributed, but centralized industrial clusters have not yet been formed, and the number of organoid innovation companies is insufficient, and it is mainly concentrated in the developed coastal areas. Enterprises with core technology advantages and complete production chains, and the earliest layout of the industry will have a first-mover advantage. With the help of policy, scientific research, capital and other factors, it is expected that more organoid companies will emerge, and the innovation ecology of the organoid industry will gradually complete.

02

类器官构建的四大要素，细胞来源：多能干细胞和成体干细胞

Organoids can be grown from Pluripotent stem cells (PSC) or Adultstem cells (ASC). PSC can be divided into Embryonic stemcell (ESC) and Induced pluripotent stemcell (iPSC). In addition, a special type of organoid is the tumor organoid, which is cultured from tumor cells isolated from biopsy samples or surgically removed tumor tissue.

（1） PSC derived organoids are good models for studying developmental processes and genetic diseases

iPSC organoids are more widely used. Human pluripotent stem cells have unlimited self-renewal and differentiate into virtually all organ-specific cell types, of which ESC has limited use due to ethical concerns. In contrast to ESC, the formation of iPSC organoids requires the reprogramming of somatic cells (usually skin or fibroblasts) into pluripotent PSCS and subsequent exposure to factors that regulate dermal development and tissue-specific stereotyping, activating or inhibiting key signaling pathways to form 3D organoids. Organoids of PSC origin can contain different cell types from different germ layers (ectoderm, mesoderm, and endoderm).

Intestinal organoids are typical endodermal derived organoids. PSC is stimulated by TGF-β signals to form a set endoderm, after which FGF4 and Wnt3A work together in a time - and dose-dependent manner to induce the formation of the posterior enteric cortex. Brain organoids are ectodermal derived organoids, and since ESCs are more similar to ectodermal cells, they can differentiate autonomously in the absence of exogenous inducers. iPSC technology enables unlimited access to specific types of cells in the human body, such as neurons, cardiomyocytes, and pancreatic beta cells, providing a rich resource for human disease modeling and drug screening. In addition, the process of PSC organoid formation follows the fate trajectory of embryonic development in vivo, so it can be used to reproduce the development process and internal changes of tissues or organs in time and space, and is a good model to study the development process and genetic diseases.

（2）ASC organoids may provide new possibilities for tissue regeneration and transplantation

Ascs are usually isolated from tissues and organs, and their ability to differentiate is very limited, and organoids can be formed in specific media containing growth factors that support their stem cell activity. The limited potency of ASC results in a single epithelial cell type in all ASC-derived organoids, lacking stromal, nervous, and vascular systems. Compared to PSC-derived organoids, the maturity of ASC-derived organoids is closer to the maturity of tissues, so they can be used to study the maintenance and regeneration of epithelial tissues, which can provide a better model for tissue repair and viral infectious diseases.

The premise of successful establishment of ASC organoids is to fully understand the regulatory stem cell niche factors and signaling pathways in vivo, and the subtle changes of each factor in the culture system may produce different culture results. For example, human Lgr5+ liver stem cells are bipotent stem cells that generate only duct epithelial organoids under standard organoid culture conditions, but have the potential to generate mature hepatocytes when Notch inhibitor, FGF19, BMP7, and dexamethasone are added to the medium.

Nowadays, a series of ASC organoids have been cultivated successively, including almost all digestive system organoids, such as liver, pancreas, colorectal, stomach, gallbladder, etc., and some non-digestive system organoids, such as prostate, breast, lung, etc. At present, human understanding of ASC is still very limited. With the in-depth study of the regulatory mechanism of ASC in vivo, ASC organoids may provide new possibilities for tissue regeneration and transplantation.

（3） Tumor organoids are the most widely used and most studied field in the field of organoids

Tumor organoids are good in vitro substitutes for patients' tumors. Tumor organoids are usually cultured from cells extracted from biopsy samples or surgically removed tumor tissue from tumor patients, and can also be cultured from fluid samples such as peripheral blood, ascites, and pleural effusion. To a large extent, tumor organoids retain the histological, pathological and genetic information of the original tumor tissue, and the tissue structure is more similar to the original tumor.

Drug susceptibility screening is one of the most common applications of tumor organoids. In 2018, the Science journal first reported on the colorectal cancer organoid drug sensitivity screening system, with a sensitivity of 100%, specificity of 93%, positive prediction rate of 88%, and negative prediction rate of 100%. More similar studies have strongly confirmed that tumor organoids derived from cancer patients can effectively predict patient clinical treatment outcomes.

At present, tumor organoids are the most widely used and the most studied field in the field of organoids. Compared with the traditional cancer cell line model and patient-derived tumor xenograft (PDX) model, tumor organoids are a model with short cycle, high efficiency and low cost, and can accurately reflect the pathophysiological state of diseases. It has great application potential in the study of cancer development, inter-tumor heterogeneity, tumor gene-drug relationship, tumor target prediction and drug sensitivity testing.

The substrate material is monopolized by Matrigel

After the cells used to build organoids are isolated, they usually need to be inoculated on biologically derived extracellular matrix materials. Extracellular matrix (ECM) is a macromolecular fiber that presents a network of gel outside the cell and provides a matrix for organoids to form three-dimensional spatial structures. Its main components include fibrin, glycoprotein, proteoglycan and glycosaminoglycan. The extracellular matrix is tissue-specific and can transmit biochemical and biomechanical signals. These signals are involved in regulating cell adhesion, migration, proliferation, and differentiation, thus guiding cell fate and development. In addition, the extracellular matrix stores and releases growth factors and other signaling molecules.

At present, the most commonly used substrate material for organoid culture is Corning's Matrigel, which has a monopoly position in the industry. Matrigel is purified from mouse sarcoma and consists mainly of laminin and collagen IV. Matrigel also has its limitations, such as high cost, batch to batch variability, and animal origin for clinical translational applications and drug detection. Therefore, it is one of the key problems that need to be solved in organoid industrialization to develop matrix adhesives with small batch difference, non-animal origin, low cost and excellent performance. 

In recent years, matrix materials have evolved from matrix gum to hydrogel systems prepared from a variety of natural or synthetic materials. Hydrogels based on natural polymers are composed of single or multiple components, proteins such as collagen and fibrin, and polysaccharides such as hyaluronic acid, alginate, glycans, and cellulose. Acellular tissue hydrogels retain important biochemical signals such as growth factors, peptides, glycan, etc., and have been used in clinical tissue regeneration, so they are also used in organoid culture research. In addition, synthetic polymer hydrogels have the advantages of molecular clarity and stable manufacturing process, thus making organoid culture more stable and repeatable. At present, the common synthetic polymer hydrogels are polyethylene glycol, polylactic-glycolic acid copolymer, polycaprolactone and so on for organoid culture.

Soluble factors guide organoid cell differentiation and proliferation

Soluble factors in the medium guide the differentiation and proliferation of organoids and promote the formation of organoids. For organoid culture, in addition to the basic medium, the participation of various soluble factors such as cytokines and small molecule inhibitors is still required. The culture conditions required for different types of organs are different, and they are highly correlated with the tissue source. Each component plays a crucial role in the culture of specific tissue-derived organoids.

The preparation of different organoids requires the use of different combinations of cytokines. The signaling pathway mediating organoid formation is the same as that of organ development and homeostasis maintenance in vivo. These soluble factors guide the differentiation and proliferation of stem cells by activating or inhibiting the signaling pathway of specific organoid types in the process of organoid culture, thus promoting the formation of organoids.

The methods of organoid culture were mainly immersed matrix glue culture

At present, organoid culture methods include submerged matrix glue culture, gas-liquid interface culture and bioreactor culture.

Submerged matrix glue culture technology is the most widely used technology in organoid culture. In this culture method, the pre-culture tissue was first made into cell suspension by mechanical shear and enzyme digestion, and then the separated and digested cells were mixed with matrix glue and inoculated into the culture plate. According to the type of organoids, specific media, cytokines and small molecule inhibitors were added for culture. Air-liquid interface culture is a highly relevant culture system in respiratory system physiology. In this method, cell suspension was obtained by tissue pretreatment, and the cell suspension was mixed with collagen gel in the cell inoculation chamber. After it solidifies, the inoculation chamber is placed in a petri dish and a specific medium is added to soak the bottom of the inoculation chamber.

The medium is diffused through a permeable membrane into the inoculation chamber to provide nutrients to the cells. Exposure of the top layer of lung cells to air can effectively promote the formation of false lamination, mucociliary differentiation and mucus secretion of lung epithelial cells, and have barrier function, so as to well simulate the in vivo characteristics of lung epithelial cells. The bioreactor culture is used for the extended culture of organoids. In the process of organoid culture, the nutrient supply and waste removal of organoids depend on the diffusion of substances, and the efficiency of material diffusion will become low when the organoids grow into larger tissue structures. Therefore, in the expanded culture of organoids, especially brain organoids, the organoids larger than millimeter can not enter in time and waste can not be removed in time, resulting in necrosis inside the organoids. The use of bioreactor culture can partly solve these problems. Bioreactor can realize real-time online monitoring of pH, temperature, dissolved oxygen, glucose and even other nutrients of the medium, and maximize nutrient supply and waste removal by means of continuous agitation and cyclic perfusion, reduce shear stress, and build expanded culture conditions suitable for organoids.

03

Organoid technology releases value, continuously expands application scenarios, and studies excellent models of developmental biology and disease

Organoid technology has been widely used in the research of developmental biology and disease modeling. Organoids are functionally and structurally similar to organs in the body and can be used to study developmental biology as well as to model disease-causing processes, including degenerative, infectious, and genetic disease models.

01 Mapping the development of the human brain using brain organs

In 2022, a Nature study showed that a scientific research team at the Swiss Federal Institute of Technology Zurich used stem cells to grow brain organoids and carefully studied their growth and development over a period of six months. Data on cell location, gene expression, and chromosome accessibility during development were studied through brain organoid technology and single-cell sequencing technology, which were then used to map human brain development to show the molecular fingerprints of every cell within the brain organoids. In 2022, a Nature study showed that a scientific research team at the Swiss Federal Institute of Technology Zurich used stem cells to grow brain organoids and carefully studied their growth and development over a period of six months. Data on cell location, gene expression, and chromosome accessibility during development were studied through brain organoid technology and single-cell sequencing technology, which were then used to map human brain development to show the molecular fingerprints of every cell within the brain organoids.

02 Brain organoids reveal human-specific disease mechanisms of neurodevelopmental diseases

Psc-derived brain organoids have been successfully used to simulate the early stages of adult brain diseases, such as Alzheimer's and Parkinson's diseases. In addition, the use of telencephalic organoids from patients with autism spectrum disorders found that the abnormal expression level of FOXG1 gene and its downstream gene resulted in excessive production of gamma-aminobutyric neuronal precursor cells and a significant increase in the number of inhibitory neuronal synapses, resulting in abnormal cortical development in children with autism spectrum disorders. This suggests that FOXG1 could be a potential drug target for idiopathic autism spectrum disorder.

03 Organoids as models for in vitro study of infectious diseases

Intestinal organoids are not only used in the pathology study of rhesus monkey rotavirus, but also as models for in vitro study of norovirus. Studies have confirmed that avian influenza H7N2 and H7N9, swine influenza H1N1, and respiratory syncytial virus can infect human bronchial organoids. In the process of fighting the novel coronavirus, lung and intestinal organoids have played an important role in studying the invasion mechanism of the novel coronavirus and screening high-throughput antiviral drugs.

Cost-effective platform for drug screening and R&D

Drugs need to be screened and evaluated before entering the clinic to determine their indication, efficacy, and safety, however, due to the limitations of existing in vitro and in vivo drug screening models, preclinical development of new drugs is slow and expensive and inefficient. The emergence of organoids provides a new cost-effective platform for drug screening and research and development, which is expected to shorten the time and improve the efficiency of new drug research and development.

01 Organoid-effect screening drugs for colorectal cancer

Hans Clevers' team used 19 colorectal cancer organoid models to screen 83 drugs, including targeted drugs for clinical application, first-line chemotherapy drugs and drugs in clinical trials, to study the relationship between drug sensitivity and PDTO molecular characteristics, and to test the effective mutation targets of clinical trial drugs. For example, IWP-2 is effective against RNF43 mutant colorectal cancer, and a potential therapeutic strategy for this mutant type has been identified.

02 Screening novel bispecific antibodies in tumor organoids

In 2022, a study in Nature Cancer was the first to screen more than 500 bispecial antibodies using an organoid bank from cancer patients and found that an EGFR × LGR5 bispecial antibody called MCLA-158 was able to effectively inhibit the growth of colorectal cancer organoids and prevent the occurrence of metastases. The study lays the groundwork for drug companies to use organoids for drug discovery.

03 Organoids are used for drug safety testing

Normal tissue organoids are used to test the safety of drugs, and cytotoxicity tests (such as lactate dehydrogenase release test) and live cell counts are used to determine the toxicity of drugs to healthy tissues. At present, a variety of healthy tissue organoids, such as liver organoids, heart organoids and kidney organoids, are used for toxicological research of drugs. Liver organoids have been used to test dose-dependent hepatotoxicity of a variety of drugs and to explore the molecular mechanisms by which some drugs cause hepatotoxicity.

A stand-in for precision medicine

At present, the use of organoids for precision medicine, especially the detection of tumor drug sensitivity, is the mainstream application scenario of the commercialization of organoids. At present, organoids can be screened for drug types including chemotherapy drugs, small molecule targeted drugs, antibody drugs and so on. The core detection indicators of the drug screen are usually IC50 and cell inhibition rate, and according to these indicators, the drug with the best tumor inhibition effect is selected among the screened drugs.

The tumor organoids are highly similar to the original patient tumors

In 2018, a Science article on the effectiveness of organoid drug susceptibility testing laid the theoretical foundation for the development of organoid drug susceptibility testing. In this study, 110 fresh tissue samples from 71 patients with colorectal cancer and gastroesophageal cancer were used to construct human organoids. It was found that the phenotype and genotype profiles of the organoids were highly similar to those of the original patients. In predicting the effectiveness of anticancer drugs, the organoids had 100% sensitivity and 93% specificity. 88% positive predictive value and 100% negative predictive value. Therefore, tumor organoids have gradually been widely used in precision medicine, providing a large-scale drug screening platform and important resources for personalized cancer treatment.

Drug susceptibility detection of tumor organoids has shown several advantages

Fast speed: high success rate of organoid construction and fast culture speed. Generally speaking, the drug screen can be performed after a week of organoid culture. The whole process from sample collection to drug sensitivity results can be well controlled within 2 weeks.

High throughput: In terms of the drug flux that can be screened, organoids can not only screen multiple drugs on the pore plate, but also test different concentrations of each drug, and multiple experiments can be carried out in parallel.

Strong clinical relevance: The clinical relevance and predictive effectiveness of organoids for cancer drug screening have been fully confirmed in a number of studies.

Regenerative medicine is on the rise

The goal of regenerative medicine is to replace or restart tissues or organs that have been damaged by disease, injury, age, or other problems. Organoids are increasingly being used as seed material and are being used in transplant studies to assess regenerative capacity. Although organoid technology is still a long way from being transformed into clinically transplantable organs, organoid transplantation offers a new hope for the research of regenerative medicine.

01 First clinical trial of organoids for refractory ulcerative colitis

On July 7, 2022, a research team at Tokyo Medical and Dental University in Japan announced the implementation of a clinical study of stem cell-derived organoids for human transplantation, and they transplanted organoids from a patient with refractory ulcerative colitis using the patient's own healthy intestinal mucosal stem cells. This attempt at regenerative medicine is the first of its kind in the world, and the safety of this trial of stem cell-derived organoids will be verified within the next year.

02 Islet organoids are used to treat diabetes

Some research teams infected iPSC cells with lentiviruses that overexpressed PD-L1, and gradually induced the production of human islet organoids. After human islet organoids were transplanted into diabetic mice with normal immune capacity, overexpressing PD-L1 was able to normalized blood sugar in the mice for more than 50 days. At present, islet organoids have successfully restored blood sugar in diabetic mice and diabetic non-human primates, and the potential of islet organoids in the treatment of diabetes is just beginning to show.

03 Organoid transplantation is used for endometrial repair and regeneration

The research team of Ningxia Medical University General Hospital successfully established mouse endometrium-derived organoids, and confirmed that the expression of these organs is highly consistent with that of mouse endometrium-derived tissues, and has stem cell characteristics and expansion ability. The mouse model of intrauterine adhesion showed that mature organoids can promote the repair and regeneration of endometrium in mice with intrauterine adhesion, and the fertility of mice with intrauterine adhesion after transplantation was obviously improved. This result fully proves that tissue damage in mice with uterine adhesions can be effectively repaired by organoid transplantation.

04

There are still many challenges ahead of the rise. Vascularization restricts the expansion of organoids and prevents the later development and maturity of organoids

Organoid technology is currently in the stage of technological explosion and scientific research results blowout, and organoid industry is also in its infancy. At present, some challenges limit the industrialization process of organoid technology, such as the vascularization problem of organoids, limited maturity and function, standardization issues, repeatability and consistency issues, and compliance issues for industrial applications.

Most organoids do not have vascularized structures of their own. With the increase in the volume and complexity of organoids, due to the restriction of oxygen and nutrient diffusion and the increase of metabolic waste, the cells inside organoids are often starved of oxygen and die, which prevents the later development and maturity of organoids and limits their clinical application.

Solution:

Addition of endothelial growth factor: Some studies have shown that in the culture of liver organoids, after supplementing vascular endothelial growth factor VEGF in the medium, vascular like structures are successfully induced in the organoids.

Co-culture: Some studies have co-cultured organoid tumor cells and vascular endothelial cells in order to generate vascular structure and solve the problem of the absence of vascularization of organoids.

Model cell growth microenvironment:The most direct way to solve the vascularization problem is to create a cell growth microenvironment in vitro that is similar to that in vivo. The primary task to solve this problem is to determine what factors play a decisive role in inducing blood vessel formation, especially to determine whether there is a limiting plate in the cell, such as the pressure of cells in different organ environments, blood composition, changes in cell pH and other factors may affect blood vessel formation.

Organoids are limited in maturity and function and cannot fully replicate human organs

Current organoid models do not fully reproduce all cell types, the level of cell maturity, and the entire physiological function of the organ, but only represent some functions of the organ. Most organoid models lack tissue-specific cell types, including immune cells, vasoforming cells, nerves, or microbiome. In addition, organoids have limited maturity, can only form foetus-like tissue rather than adult tissue, and lack important physiological processes in the body as well as blood vessels, lymphatic vessels and nerve functions, and are still unable to fully replicate human organs.

Solution:

Use of small molecule compounds: In order to overcome the problem of organoid maturity and limited function, many programs in the research process include the use of small molecule compounds (such as BDNF) to pretreat the organoids to accelerate the maturation of the organoids.

Bioreactor: The use of bioreactor to culture organoids can partially solve the oxygen and nutrient needs of organoids and promote the growth and maturity of organoids.

Animal chimera technology: In addition, xenotransplantation forms organoids - Animal chimera technology can promote organoids to form nerve or blood vessel networks.

The organs of the same kind cultivated by different programs are different and lack of standardization

At present, multiple tissues and organs and multiple tumor organoids have been successfully cultured, and the cultivation of organoids is no longer the primary problem. However, the organoid culture technology and medium scheme adopted by different laboratories and companies are different, especially the soluble factors such as growth factors, amino acids and small molecule inhibitors used are different, resulting in the difference of organoids cultured by different schemes, and the experimental results obtained by different application scenarios will vary to different degrees. Organoids are facing standardization problems from laboratory research to industrial application.

Solution:

Expert consensus: At present, both international and domestic are actively promoting the standardization of organoids, and forming expert consensus in different application scenarios such as quality control standards and clinical applications.

Develop standards for the whole chain of organoids: The organoid industry still needs stronger documents to promote the standardization of the industry, and develop standards and parameters for the entire chain of organoids preparation, whether it is the transportation of clinical samples, the quality standards of clinical samples, the preparation standards and culture standards of organoids, as well as the operational processes and key quality control points of application scenarios.

Repeatability and consistency are major bottlenecks in the commercialization of organoid technologies.

In the process of organoid culture, drug screening and drug sensitivity screening, manual operation mode is still used, and human factors are too involved and the degree of automation is low, resulting in large errors caused by systematic chance, and poor repeatability and consistency of experimental results, which is difficult to meet the needs of commercial applications and limits the transformation of organoids into clinical research and commercial applications.

Solution:

Automation System: In order to promote the development and industrial application of this cutting-edge technology, Bozhen Biology combined with the actual pain points in the development and application of organoid technology, and made every effort to create the world's first automatic culture and high-throughput drug screening system for organoids, solving the urgent problems of high cost, low throughput, poor data consistency and repeatability of organoids at this stage. It can realize one-stop line production of organoids from primary culture, model construction to drug evaluation, and realize standardized, intelligent and automated operation of the whole process.

Compliance development is a long-term strategy for the development of organoid industry

The high-quality industrial development of the organoid industry must open up the compliance path from the collection and collection of human genetic resources such as tissues and cells to the cultivation and construction of the library of organoids and the final industrial application process of human genetic resources supervision, biosafety, personal information protection and data security. With the implementation of the regulations on the management of human genetic resources, the national requirements for the use of human genetic resources are becoming more and more standardized, and how to obtain and use organoids and primary cells legally and in compliance is a problem that must be considered.

Solution:

Definition of laws and regulations: On the issue of the industrial application of organoid technology, the state has yet to make further and more detailed provisions at the legal and regulatory levels.

Cooperation with the national human genetic resources Bank: It is a worthwhile path for organoid commercial companies to discuss cooperation with the national human genetic resources bank at the strategic and business levels. Through joint investment, they can build organoid sample banks, develop organoid models, and achieve legal and compliant industrial applications.

05

Future outlook: Organochip integrates the advantages of organochip and organochip, and expands the application boundary

Despite the great potential of organoids, existing organoid culture systems have some limitations that limit their wide application. As a result, researchers are beginning to take inspiration from the fields of bioengineering and materials to produce organoids that are more physiologically relevant and suitable for target applications. Great advances in micro-nano processing technology and microfluidic technology have made remarkable progress in organ chips. Organ-on-a-chip refers to the human organ physiological microsystem composed of cells, extracellular matrix, sensors to detect cell function, and other elements with microfluidic technology as the core. The main advantage is that it can precisely control cells and their microenvironment.

Based on this, organoid-on-a-chip integrates the advantages and characteristics of organoids and organ chips, and integrates a variety of functional structural units, such as Organoid culture chambers, microfluidics, actuators, biosensors, etc., to form an organ physiological microsystem. Organoid chips not only have the advantages of organoids, which can simulate the development process, physiological state and function of organs, and recombine the functions and physiological structure of source tissues, but also have the advantages of organ chips that can accurately control cells and their microenvironment, realizing the control of fluid at the micron scale and dynamic capture of parameter changes. The simulation degree and sensitivity of the biological model to the change of experimental parameters are further improved.

Organochip integrates the advantages of organoids and organochips to expand the application boundary

In recent years, organoid chip technology has made some important progress, and researchers have successively established multi-type organ chip systems, such as brain, intestine, liver, islet, kidney, retina, etc., as well as organoid interaction chips that integrate multiple types of organs. The organoid chip is characterized by micro-structure, high throughput and high sensitivity, and integrates a series of experimental processes such as sorting, culture, observation, stimulation induction, detection and analysis of organoids into one, which has been initially applied in developmental biology, drug testing, disease modeling and other fields.

Sum up

Organochip integrates the advantages of organoids and organochips to expand the application boundary

Organochip technology is conducive to organoid flux generation and vascularization formation, which enables organoids to have higher repeatability and more complex structure, and can simulate the interaction conditions of multiple types of organs and the control of tissue microenvironment, partially making up for the limitations of the existing organoid system, and becoming one of the future development directions of organoid technology. 

Organoid technology is integrated with a variety of innovative technologies to expand the application boundary. Organoid technology, combined with live cell imaging and high-throughput analysis technology, can track the formation and development of organoids in real time, and carry out high-throughput drug screening and evaluation. Combined with 3D bioprinting and new biomaterials, functional organoids with multilevel complex structure and large scale can be prepared. Combined with gene editing, multi-omics analysis, artificial intelligence and other technologies, it can further improve the accuracy of organoids to reflect human physiological or pathological processes, and help to deeply understand and deeply analyze the development process and disease mechanism of organs with higher fidelity. This will require the collaboration of many interdisciplinary researchers to promote the development and transformation of organoid technology.

Organoid intelligence opens a new paradigm of biological computing based on brain-like organs

The human brain is arguably the most powerful computing system known. It is highly efficient at processing large amounts of complex information, can distinguish signals from noise, adapt and filter error messages, all while running on just 20 watts of power. The processing efficiency, progressive learning and plasticity of the human brain are unmatched by any computer system. The human brain is better at processing small or uncertain amounts of data; Sequential processing can be performed, or parallel processing can be performed, while computers can only perform sequential processing; Human brains outperform computers when it comes to making decisions with large, highly heterogeneous, incomplete data sets and other challenging forms of processing.

Based on the powerful computational energy of the human brain and the great potential of brain organoid research, Johns Hopkins University in the United States organized the first organoid Intelligence seminar in 2022. At the meeting, a new term and emerging field - organoid intelligence was creatively proposed, aiming to establish organoid intelligence as a true form of biological computing and release the great potential of brain organoids in biological intelligence. This led to the formation of an organoid intelligence research community and the adoption of the Baltimore Declaration for the Exploration of Organoid Intelligence.

Professor Thomas Hartung, one of the leaders of organoid intelligence research, demonstrated a biocomputer system based on organoid intelligence. At the heart of the system are 3D human brain organs that perform calculations. The learning potential of organoids is optimized through culture conditions and enrichment of cells and genes that are critical for learning. The scalability, feasibility and durability of organoids are supported by an integrated microfluidic system. Various types of inputs can be provided to organoids, including electrical and chemical signals, synthetic signals from machine sensors, and natural signals from connected sensory organs such as the retina. High-resolution output measurements are made with electrophysiological recordings obtained by microelectric arrays and implantable probes, as well as imaging of organoid structures and functional properties. These outputs can be used directly for computation and as biofeedback to facilitate organoid learning. Artificial intelligence and machine learning are used to encode and decode signals and develop hybrid biological computing solutions in conjunction with suitable big data management systems.

At present, organoid intelligence is still in the stage of concept proposal and preliminary exploration, and the realization of biological computer systems is very challenging, and there are many technical problems to be solved and breakthroughs. For example, bioengineering technology (induced pluripotent stem cell technology) and organoid culture technology are used for brain organoid culture; Microfluidic technology simulates the structure and culture environment of real brain to support the homeostasis and viability of organoids. 3D microelectrode arrays and implantable electrophysiological devices for high-resolution electrophysiological signal recording; Complex, diverse and controllable signal inputs; Ai correlates and interprets the input and output of organoid intelligent systems; Big data storage infrastructure for organoid intelligent systems.

In addition to the above-mentioned biological computer systems for computing, learning and storing memories, organoid intelligence can also be used to explore the specific processes of neurodevelopment and the specific causes of neurodegenerative diseases (such as Alzheimer's disease), neurodevelopmental disorders (such as autism), schizophrenia and other neurological diseases, and guide the development and application of new treatment options.

The concept of organoid intelligence will trigger a new revolution in the field of biological computing, open a new paradigm of biological computing based on brain-like organs, overcome the limitations of computer-based artificial intelligence, and have a significant impact on the world. Specifically, biocomputing based on organoid intelligence allows for faster decision making (including decisions on large, incomplete, and heterogeneous data sets), continuous learning during tasks, and greater energy utilization and data storage efficiency. In addition, the development of organoid intelligence will also provide tremendous opportunities to elucidate the biological basis of human cognition, learning, and memory, as well as various diseases associated with cognitive deficits, helping to identify new therapeutic approaches that could address numerous unmet clinical needs.