Regenerative Medicine Research Articles

A Tympanic Piezo-Bioreactor Modulates Ion Channel-Associated Mechanosignaling to Stabilize Phenotype and Promote Tenogenesis in Human Tendon-Derived Cells.

Preserving the function of human tendon-derived cells (hTDCs) during cell expansion is a significant challenge in regenerative medicine. In this study, a non-genetic approach is introduced to control the differentiation of hTDCs using a newly developed tympanic bioreactor. The system mimics the functionality of the human tympanic membrane, employing a piezoelectrically tuned acoustic diaphragm made of polyvinylidene fluoride-co-trifluoroethylene and boron nitride nanotubes. The diaphragm is vibrationally actuated to deliver targeted electromechanical stimulation to hTDCs. The results demonstrate that the system effectively maintains the tendon-specific phenotype of hTDCs, even under conditions that typically induce nonspecific differentiation, such as osteogenesis. This stabilization is achieved by modulating integrin-mediated mechanosignaling via ion channel-regulated calcium activity, potentially by TREK-1 and PIEZO1, yet targeted studies are required for confirmation. Finally, the system sustains the activation of key differentiation pathways (bone morphogenetic protein, BMP) while downregulating osteogenesis-associated (mitogen-ctivated protein kinase, MAPK and wingless integrated, WNT) pathways and upregulating Focal Adhesion Kinase (FAK) signaling. This approach offers a finely tunable, dose-dependent control over hTDC differentiation, presenting significant potential for non-genetic approaches in cell therapy, tendon tissue engineering, and the regeneration of other mechanosensitive tissues.

PERSPECTIVES FOR THE USE OF PLANT RAW MATERIALS OF THE CLOVER MEADOW (TRIFOLIUM PRATENSE L.) IN PHARMACY

Background. The creation and registration of original medicines based on plant raw materials are becoming promising areas in pharmaceutical practice. Clover meadow (Trifolium pratense L.) has been used in folk medicine for a long time, due to the content of many biologically active substances with Therapeutic properties. It grows over large areas; extracts are easy to obtain from plant raw materials and have a certain economic attractiveness. In this regard, compounds extracted from meadow clover can be considered as potential precursors for the development of new promising pharmaceuticals. Aim. Carrying out a narrative review on the Perspectives of the use of biologically active substances extracted from the plant raw materials of meadow clover in pharmaceutical practice. Material and methods. The scientific publications of the PubMed biomedical research database are analyzed. Results. Meadow clover (Trifolium pratense L.) is a widespread perennial herbaceous plant belonging to the pharmacopoeia group. Extracts obtained from plant raw materials have a wide range of biological activity. The largest proportion of them are isoflavones, flavonoids, saponins, clovamides and phenolic acids. Isoflavones have a phytoestrogenic effect, and in this regard, they are successfully used in the treatment of diseases of the reproductive system. In addition, their anti-inflammatory, reparative properties have been proven; they can be used for the prevention and recovery of metabolic disorders. Flavonoids, saponins, clovamides and phenolic acids have antioxidant and antiplatelet effects. Conclusion. Among biologically active substances extracted from meadow clover, the greatest use in pharmacological practice was noted for isoflavones. At the same time, there are a few other potentially effective compounds with known medicinal properties. Their further study is required in order to create new original medicines

Functionality of lyophilized osteoinductive EVs: a mechanistic study

IntroductionMesenchymal stem cell-derived extracellular vesicles (MSC EVs) hold significant promise for regenerative medicine. Lyophilization of EVs significantly enhances their translational potential. While, lyophilized EVs have been studied from a morphological perspective, the functional stability of these EVs and their cargo following lyophilization need to be mechanistically investigated.MethodsIn this study, we investigated the functional and mechanistic bioactivity of fresh versus lyophilized MSC EVs, specifically focusing on functionally engineered osteoinductive EVs developed in our laboratory. We utilized dimethyl sulfoxide (DMSO) as a cryoprotectant and conducted pathway-specific in vitro and in vivo experiments to assess the stability and functionality of the EVs.ResultsOur findings show that using DMSO as a cryoprotectant before lyophilization preserves the functional stability of engineered MSC EVs. In vitro experiments demonstrated that the endocytosis, cargo integrity, and pathway-specific activity of lyophilized EVs were maintained when DMSO was used as the cryoprotectant. Additionally, in vivo bone regeneration studies revealed that the functionality of cryoprotected lyophilized EVs was comparable to that of freshly isolated EVs.DiscussionThese results provide a foundation for evaluating the functionality of lyophilized EVs and exploring the use of DMSO and other cryoprotectants in EV-based therapies. Understanding the functionality of lyophilized naïve and engineered EVs from a mechanistic perspective may enhance validation approaches for tissue regeneration strategies.

Perspectives on non-genetic optoelectronic modulation biointerfaces for advancing healthcare

AbstractAdvancements in optoelectronic biointerfaces have revolutionized healthcare by enabling targeted stimulation and monitoring of cells, tissues, and organs. Photostimulation, a key application, offers precise control over biological processes, surpassing traditional modulation methods with increased spatial resolution and reduced invasiveness. This perspective highlights three approaches in non-genetic optoelectronic photostimulation: nanostructured phototransducers for cellular stimulation, micropatterned photoelectrode arrays for tissue stimulation, and thin-film flexible photoelectrodes for multiscale stimulation. Nanostructured phototransducers provide localized stimulation at the cellular or subcellular level, facilitating cellular therapy and regenerative medicine. Micropatterned photoelectrode arrays offer precise tissue stimulation, critical for targeted therapeutic interventions. Thin-film flexible photoelectrodes combine flexibility and biocompatibility for scalable medical applications. Beyond neuromodulation, optoelectronic biointerfaces hold promise in cardiology, oncology, wound healing, and endocrine and respiratory therapies. Future directions include integrating these devices with advanced imaging and feedback systems, developing wireless and biocompatible devices for long-term use, and creating multifunctional devices that combine photostimulation with other therapies. The integration of light and electronics through these biointerfaces paves the way for innovative, less invasive, and more accurate medical treatments, promising a transformative impact on patient care across various medical fields.

Fabrication and development of mechanical metamaterials via additive manufacturing for biomedical applications: A review

Abstract In this review, we provide a comprehensive overview of additive manufacturing (AM) technologies and materials as well as design possibilities to manufacture metamaterials for a variety of biomedical applications, of which many are inspired by nature itself. It is described that how new AM technologies (such as continuous liquid interface production and multiphoton polymerization) and new developments in more mature AM technologies (such as powder bed fusion, stereolithography and extrusion-based bioprinting) have led to more precise, efficient, and personalized biomedical components. Extrusion-based bioprinting, specifically, is a revolutionary topic creating intricate models with remarkable mechanical compatibility of metamaterials, such as negative/zero Poisson's ratio and stress elimination for tissue engineering and regenerative medicine. By exploiting a variety of designs for porous structures (such as truss, triply periodic minimal surface, and plant/animal-inspired and functionally graded lattices), AM made bioactive bone implants, artificial tissues and organs are made for tissue replacement purposes. The material palette of the AM metamaterials has also become very diverse nowadays ranging from metals and alloys (such as titanium and cobalt-chromium alloys) to polymers (like biodegradable polycaprolactone (PCL) and polymethyl methacrylate (PMMA)) which could be even integrated within bioactive bioceramics. These advancements are driving the progress of biomedical technology, improving human health and quality of life for future generations.

Lactobacillus rhamnosus GG-derived extracellular vesicles promote wound healing via miR-21-5p-mediated re-epithelization and angiogenesis

Extracellular vesicles (EVs), especially those derived from stem cells, have emerged as a novel treatment for promoting wound healing in regenerative medicine. However, the clinical application of mammalian cells-derived EVs is hindered by their high cost and low yields. Inspired by the ability of EVs to mediate interkingdom communication, we explored the therapeutic potential of EVs released by the probiotic strain Lactobacillus rhamnosus GG (LGG) in skin wound healing and elucidated the underlying mechanism involved. Using full-thickness skin wound-healing mouse models, we found that LGG-EVs accelerated wound healing procedures, including increased re-epithelialization and promoted angiogenesis. Using in vitro experiments, we further demonstrated that LGG-EVs boosted the proliferation and migration capacities of both epithelial and endothelial cells, as well as promoted endothelial tube formation. miRNA profiling analysis revealed that miR-21-5p was highly enriched in LGG-EVs and LGG-EV treatment significantly increased miR-21-5p level in recipient cells. Mechanically, LGG-EVs induced regulatory effects via miR-21-5p mediated metabolic signaling rewiring. Our results suggest that EVs derived from LGG could serve as a promising candidate for accelerating wound healing and possibly for treating chronic and impaired healing conditions.Graphical abstract

Natural polymeric biomaterials for managing peripheral nerve injuries : a novel approach for tissue repair and reconstruction.

Peripheral nerve injury serves as a major challenge to clinicians and researchers due to the complexity and functions of peripheral nerves that play the crucial role of transmitting signals between spinal cord and other human body tissues. Biomaterials offer promising solutions for regeneration of nerve tissues owing to their biodegradability, and biocompatibility. They can be fabricated as hydrogels, scaffolds, nanofibrous matrices, and nanoparticles that can facilitate the release of therapeutic molecules to reduce inflammation and promote neuronal growth. Managing peripheral nerve injuries through natural polymeric based biomaterials represents as a novel approach for tissue repair and reconstruction in the field of regenerative medicine. This correspondence will give an insight into the different types of natural polymeric biomaterials that can be efficiently used in managing peripheral nerve injuries.

Advances in Cell Therapy Using Rabbit Models for Regenerative Medicine

Cell therapy represents an innovative and promising approach to treating debilitating conditions, and animal testing on rabbits has played a crucial role in advancing this therapy. The objective of this manuscript aims to review the literature on the use of lagomorphs in cell therapy studies. The Pubmed and ScienceDirect databases were used, with the descriptors: Cell therapy and experimentation in rabbits; rabbits and cell therapy. After an initial retrieval of 1799 articles, those that did not specifically address the use of rabbits in cell therapy or were unrelated to the scope of this review were excluded. This resulted in a final selection of 21 articles. The present work presents a broad and distinct view of cell therapy and its different applicability, presenting a promising framework for animal experimentation in rabbits, and the advantages of using this animal model.

Use of Advanced Platelet-Rich Fibrin for the Treatment of Multiple Adjacent Mucogingival Recessions: A Technical Report of the Fibrin-Assisted Soft-Tissue Promotion Protocol.

Platelet-rich fibrin (PRF) has had a marked impact on regenerative medicine due to its widespread ability to promote angiogenesis to defective tissues. Particularly in the dental field, evidence from randomized clinical trials has further shown that PRF facilitates greater soft-tissue regeneration when compared with hard tissues. Recently, the fibrin-assisted soft-tissue promotion (FASTP) technique has been developed as a means to promote soft-tissue regeneration of mucogingival recessions utilizing PRF. Within the present case report, a 28-year-old male presented with multiple adjacent mucogingival recessions in the maxilla ranging in probing depths (1-3 mm) and gingival recessions (1-5 mm). For optimal regenerative outcomes, the use of advanced PRF (A-PRF; 1300 RPM for 8 min) has been utilized to enhance regenerative outcomes by fully taking advantage of the low-speed centrifugation concept (LSCC). This case report highlights the latest surgical concepts, centrifugation protocols, and use of the LSCC to regenerate multiple adjacent mucogingival recessions in the esthetic zone.

The role of biophysical cues and their modulated exosomes in dental diseases: from mechanism to therapy.

Dental diseases such as caries and periodontitis have been common public health problems. Dental disease treatment can be achieved through stem cell-based dental regeneration. Biophysical cues determine the fate of stem cells and govern the success of dental regeneration. Some studies have manifested exosomes derived from stem cells could not only inherit biophysical signals in microenvironment but also evade some issues in the treatment with stem cells. Nowadays, biophysical cue-regulated exosomes become another promising therapy in dental regenerative medicine. However, methods to improve the efficacy of exosome therapy and the underlying mechanisms are still unresolved. In this review, the association between biophysical cues and dental diseases was summarized. We retrospected the role of exosomes regulated by biophysical cues in curing dental diseases and promoting dental regeneration. Our research also delved into the mechanisms by which biophysical cues control the biogenesis, release, and uptake of exosomes, as well as potential methods to enhance the effectiveness of exosomes. The aim of this review was to underscore the important place biophysical cue-regulated exosomes occupy in the realm of dentistry, and to explore novel targets for dental diseases.

Generation of IPi002-A/B/C human induced pluripotent stem cell lines from MARCH amniotic fluid cells

Human induced pluripotent stem cells (hiPSCs) have become a revolutionary tool in biomedical research due to their unique in vitro properties and fate versatility. They offer insights into development or genetic disorders, facilitate drug discovery and hold promise for regenerative medicine. Here we generated three hiPSC cells – IPi002-A/B/C – from primary amniotic fluid cells (AFCs) obtained via amniocentesis for the prenatal diagnosis of MARCH syndrome: Multinucleated neurons, Anhydramnios, Renal dysplasia, Cerebellar hypoplasia, and Hydranencephaly. These AFCs underwent reprogramming through non-integrative viral transduction and the resulting hiPSCs exhibited normal karyotype and expressed typical pluripotency markers.

Innovative Biomedical Interventions for Cardiovascular Risk Reduction and Health Promotion: A Review Study

Cardiovascular diseases (CVD) remain the leading cause of mortality worldwide, with increasing prevalence in both developed and developing regions. The complexity of CVD necessitates innovative approaches that combine biomedical research with clinical practice for effective prevention, diagnosis, and treatment. This chapter explores cutting-edge biomedical interventions to reduce cardiovascular risk and promote heart health. It focuses on recent advancements in biomarker identification, gene therapy, nanotechnology, and regenerative medicine, offering new strategies for early detection and targeted treatment of cardiovascular conditions. Additionally, the chapter highlights the role of precision medicine and personalized healthcare approaches in managing individual risk factors, including hypertension, dyslipidemia, and diabetes, while addressing lifestyle influences like nutrition and physical activity. Public health initiatives are also discussed, examining how biomedical research has contributed to the development of population-level interventions, such as community-based screening programs and digital health tools for monitoring cardiovascular health. By integrating biomedical innovation with preventive healthcare strategies, this chapter provides a comprehensive overview of the future directions in cardiovascular risk reduction, underscoring the importance of a multi-disciplinary approach for enhancing patient outcomes and promoting long-term cardiovascular wellness.

Sustainable bioinspired materials for regenerative medicine: balancing toxicology, environmental impact, and ethical considerations

The pursuit of sustainable bioinspired materials for regenerative medicine demands a nuanced balance between scientific advancement, ethical considerations, and environmental consciousness. This abstract encapsulates a comprehensive perspective paper exploring the intricate dynamics of toxicology, environmental impact, and ethical concerns within the realm of bioinspired materials. As the landscape of regenerative medicine evolves, ensuring the biocompatibility and safety of these materials emerges as a pivotal challenge. Our paper delves into the multidimensional aspects of toxicity assessment, encompassing cytotoxicity, genotoxicity, and immunotoxicity analyses. Additionally, we shed light on the complexities of evaluating the environmental impact of bioinspired materials, discussing methodologies such as life cycle assessment, biodegradability testing, and sustainable design approaches. Amid these scientific endeavors, we emphasize the paramount importance of ethical considerations in bioinspired material development, navigating the intricate web of international regulations and ethical frameworks guiding medical materials. Furthermore, our abstract underscores the envisioned future directions and challenges in toxicology techniques, computational modeling, and holistic evaluation, aiming for a comprehensive understanding of the synergistic interplay between sustainable bioinspired materials, toxicity assessment, environmental stewardship, and ethical deliberation.

Wnt specifically induces FZD5/8 endocytosis and degradation and the involvement of RSPO-ZNRF3/RNF43 and DVL.

Frizzled (FZD) proteins are the principal receptors of the Wnt signaling pathway. However, whether Wnt ligands induce FZD endocytosis and degradation remains elusive. The transmembrane E3 ubiquitin ligases ZNRF3 and RNF43 promote the endocytosis and degradation of FZD receptors to inhibit Wnt signaling, and their function is antagonized by R-spondin (RSPO) proteins. However, the dependency of RSPO-ZNRF3/RNF43-mediated FZD endocytosis and degradation on Wnt stimulation, as well as the specificity of this degradation for different FZD, remains unclear. Here, we demonstrated that Wnt specifically induces FZD5/8 endocytosis and degradation in a ZNRF3/RNF43-dependent manner. ZNRF3/RNF43 selectively targets FZD5/8 for degradation upon Wnt stimulation. RSPO1 enhances Wnt signaling by specifically stabilizing FZD5/8. Wnt promotes the interaction between FZD5 and RNF43. We further demonstrated that DVL proteins promote ligand-independent endocytosis of FZD but are dispensable for Wnt-induced FZD5/8 endocytosis and degradation. Our results reveal a novel negative regulatory mechanism of Wnt signaling at the receptor level and illuminate the mechanism by which RSPO-ZNRF3/RNF43 regulates Wnt signaling, which may provide new insights into regenerative medicine and cancer therapy.

Multispectral Imaging of Collagen, NAD(P)H and Flavin Autofluorescence in Mesenchymal Stem Cells Undergoing Trilineage Differentiation.

Understanding the molecular mechanisms of differentiation is important for regenerative medicine and developmental biology. This study aims to characterise the role of the glycolysis/oxidative phosphorylation balance as a driver of mesenchymal stem cell (MSC) differentiation. Cells were maintained in normal conditions or stimulated towards the MSC trilineage cell types over 21 days. Multispectral imaging of cell autofluorescence was applied as a non-invasive methodology to continuously image cultures in situ. Spectral signals for collagen, NAD(P)H, and flavins were unmixed. MSCs cultured under chondrogenic conditions exhibited increased collagen levels relative to controls. Following osteogenic induction, MSCs showed increased collagen levels relative to controls during the earlier stages of culture; however, control cells increased their collagen levels as they became confluent. MSCs cultured under adipogenic conditions exhibited lower levels of collagen than controls. The redox ratio (RR; NAD(P)H/flavins) immediately decreased during chondrogenesis, with this early effect persisting throughout the culture compared to control cells, which appeared to increase their RR, similar to osteogenesis. Adipogenesis resulted in a small increase in RR on day 2 relative to control cells, followed by a persistent decrease. Chondrogenic and adipogenic differentiation favoured oxidative phosphorylation, whereas osteogenesis and MSC overgrowth resulted in a glycolytic metabolism. Following consideration of these findings, as well as the diverse reports in the literature, it is concluded that neither enhanced oxidative phosphorylation nor glycolysis are fundamental to the canonical modes of differentiation, and researchers should avoid interpreting shifts as indicating differentiation.

3D Bioprinting for Engineered Tissue Constructs and Patient-Specific Models: Current Progress and Prospects in Clinical Applications.

Advancements in bioprinting technology are driving the creation of complex, functional tissue constructs for use in tissue engineering and regenerative medicine. Various methods, including extrusion, jetting, and light-based bioprinting, have their unique advantages and drawbacks. Over the years, researchers and industry leaders have made significant progress in enhancing bioprinting techniques and materials, resulting in the production of increasingly sophisticated tissue constructs. Despite this progress, challenges still need to be addressed in achieving clinically relevant, human-scale tissue constructs, presenting a hurdle to widespread clinical translation. However, with ongoing interdisciplinary research and collaboration, the field is rapidly evolving and holds promise for personalized medical interventions. Continued development and refinement of bioprinting technologies have the potential to address complex medical needs, enabling the development of functional, transplantable tissues and organs, as well as advanced in vitro tissue models.

Maternal and neonatal factors’ effects on wharton's jelly mesenchymal stem cell yield

As Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) are easily accessible, easy to isolate, and ethically acceptable, they represent a promising source of MSCs for use in regenerative medicine. Considering decisions on WJ-MSCs collection requires extensive knowledge of the factors that impact their yield. This study's aim was to evaluate the influence of parameters related to mothers and newborns on the WJ-MSCs yield. The WJ-MSCs were isolated and expanded after being isolated from 79 umbilical cord (UC) samples. Population doubling time and cell proliferation were assessed. By flow cytometry analysis, WJ-MSCs were identified by positivity of CD105, CD90, and CD73 and negativity of CD45 and CD34. There was a statistically significant negative correlation between UC width and P1 doubling time. Maternal age and WJ-MSC yield were shown to be negatively correlated. Birth weight and gestational age showed a significant positive correlation between WJ-MSCs yield and neonatal variables. No significant correlations were detected between the WJ-MSCs and the mother parity, nor the neonatal sex, fetal presentation, or head circumference. The WJ-MSCs yield increases with younger maternal age, higher gestational age, and increased neonatal birth weight. Hence, consideration should be given to these factors when selecting the ideal donors.

Hydrogels Innovation: A Review on Recent Development, Characterization, and Applications

Hydrogels are special materials that can hold a large amount of water and form 3D networks. In the past few years, there have been exciting improvements in hydrogel technology, bringing new ideas to many different areas. By trying out new materials like smart hydrogels that can respond to different conditions, provided new ways to deliver medicine precisely, build tissues, and create wearable gadgets. Among the most significant developments is the creation of smart hydrogels, which can react dynamically to different environmental stimuli. With their ability to release therapeutic chemicals under regulated conditions in response to particular physiological cues, these intelligent materials have enormous potential for the administration of precision medicine. These kinds of customized drug delivery systems have the power to completely change how treatments are administered by reducing adverse effects and increasing therapeutic efficacy. Hydrogels are also useful in tissue engineering, where they are used as scaffolds to create biological tissues that function. Hydrogel-based tissue constructions, which imitate the extracellular matrix, offer a favorable microenvironment for cell proliferation and differentiation, promoting the healing of injured or ill tissues. With its enormous potential in regenerative medicine, this revolutionary strategy offers hope for the treatment of ailments including organ failure. this article gives a thorough look at the recent developments in hydrogels, characterization techniques, and the new application of hydrogels in various fields of science. In summary, new developments in hydrogel technology have opened up a plethora of opportunities in a variety of scientific fields. The adaptable properties of hydrogels continue to spur innovation in a variety of fields, including wearable technology, tissue engineering, and precision medicine. These applications offer revolutionary answers to urgent social issues.

Transcriptome-Optimized Hydrogel Design of a Stem Cell Niche for Enhanced Tendon Regeneration.

Bioactive hydrogels have emerged as promising artificial niches for enhancing stem cell-mediated tendon repair. However, a substantial knowledge gap remains regarding the optimal combination of niche features for targeted cellular responses, which often leads to lengthy development cycles and uncontrolled healing outcomes. To address this critical gap, an innovative, data-driven materiomics strategy is developed. This approach is based on in-house RNA-seq data that integrates bioinformatics and mathematical modeling, which is a significant departure from traditional trial-and-error methods. It aims to provide both mechanistic insights and quantitative assessments and predictions of the tenogenic effects of adipose-derived stem cells induced by systematically modulated features of a tendon-mimetic hydrogel (TenoGel). The knowledge generated has enabled a rational approach for TenoGel design, addressing key considerations, such as tendon extracellular matrix concentration, uniaxial tensile loading, and in vitro pre-conditioning duration. Remarkably, our optimized TenoGel demonstrated robust tenogenesis in vitro and facilitated tendon regeneration while preventing undesired ectopic ossification in a rat tendon injury model. These findings shed light on the importance of tailoring hydrogel features for efficient tendon repair. They also highlight the tremendous potential of the innovative materiomics strategy as a powerful predictive and assessment tool in biomaterial development for regenerative medicine.

Potential Use of Organoids in Regenerative Medicine.

In vitro cell culture is crucial for studying human diseases and development. Compared to traditional monolayer cultures, 3D culturing with organoids offers significant advantages by more accurately replicating natural tissues' structural and functional features. This advancement enhances disease modeling, drug testing, and regenerative medicine applications. Organoids, derived from stem cells, mimic tissue physiology in a more relevant manner. Despite their promise, the clinical use of regenerative medicine currently needs to be improved by reproducibility, scalability, and maturation issues. This article overviews recent organoid research, focusing on their types, sources, 3D culturing methods, and applications in regenerative medicine. A literature review of "organoid" and "regenerative medicine" in PubMed/MEDLINE highlighted relevant studies published over the past decade, emphasizing human-sourced organoids and their regenerative benefits, as well as the availability of free full-text articles. The review uses descriptive data, including tables and text, to illustrate the challenges and potential of organoids in regenerative medicine. The transition from 2D to 3D models, particularly organoids, has significantly advanced in vitro research. This review covers a decade of progress in various organoid types-such as liver, cholangiocyte, intestinal, pancreatic, cardiac, brain, thymus, and mammary organoids-and their 3D culture methods and applications. It addresses critical issues of maturity, scalability, and reproducibility and underscores the need for standardization and improved production techniques to facilitate broader clinical applications in regenerative medicine. Successful therapy requires increased scalability and standardization. Organoids have enormous potential in biological research, notwithstanding obstacles.