Sci. Chung, C., Lampe, K. J. Biomater. USA 113, E6831E6839 (2016). Mater. Google Scholar. Vigneswaran, Y. et al. A droplet microfluidic system to fabricate hybrid capsules enabling stem cell organoid engineering. Amikagels with elastic moduli ranging from 37 to 320kPa were created, but no chemical signaling peptides were incorporated into the gels. Other blood vessels are structurally intact, despite cell removal (e), with the structural details apparent even at high magnification (f). 11, 3550 (2014). found that decellularized human ECM is capable of supporting the survival of undifferentiated hPSCs and their pancreatic lineage derivatives, including insulin-expressing beta-like cells, in vitro81. The authors also established that the stiffness of this hybrid material is relevant to cell fate determination; lower elastic moduli, ca. J. Respir. Curr. Designer matrices for intestinal stem cell and organoid culture. Recombinant proteins made by genetically engineered organisms have found wide applications in medicine, food processing, and catalysis. 6, 402420 (2021). hPSC lines should be validated for pluripotency and normal karyotype prior to use. Artificial niche microarrays for probing single stem cell fate in high throughput. Decellularized ECM is also chemically undefined; the factors driving differentiation are often unknown. Li, N. K., Quiroz, F. G., Hall, C. K., Chilkoti, A. Sci. Hong, X. et al. 5). Synthetic hydrogels can be functionalized with biologically active moieties that permit the growth and spread of cells146,147,148,149,150,151; concentration and spacing of these cues can be changed independently152,153,154,155. Carbohydr. The elastic modulus of matrigel (TM) as determined by atomic force microscopy. Differential effects of heparin and hyaluronic acid on neural patterning of human induced pluripotent stem cells. The generation of cancer organoids from. Rep. 7, 11 (2017). Commun. Endometrial Assembloids to Model Human Embryo Implantation - Springer & Jiang, P. Three-dimensional hyaluronic acid hydrogel-based models for in vitro human iPSC-derived NPC culture and differentiation. Drost, J. A functional CFTR assay using primary cystic fibrosis intestinal organoids. Choi, J.-W. et al. Wylie, R. G. et al. Anderson, D. G., Levenberg, S. & Langer, R. Nanoliter-scale synthesis of arrayed biomaterials and application to human embryonic stem cells. Immunol. Ed. Hof, K. S. & Bastings, M. M. C. Programmable control in extracellular matrix-mimicking polymer hydrogels. 18, 573 (2017). Choi, H., Song, J., Park, G. & Kim, J. The authors used a 4-arm PEG maleimide to encapsulate and culture Matrigel-derived HIOs. Cell Sci. 167, 216219 (2009). Further information on research design is available in theNature Research Reporting Summary linked to this article. Rossen, N. S. et al. 4, a004903a004903 (2012). Med. J. Mol. This method has the advantage of allowing easy harvesting of cells, which are simply allowed to settle to the bottom of a tube. Stem Cells Int. Sci. Further work should be done in this system to establish long-term viability. & Chang, D. T. Foreign body reaction to biomaterials. Contrarily, myocardial infarct is known to trigger remodeling events that stiffen the ECM and change its chemical composition; yet when mesenchymal stem cells are seeded on infarcted tissue, the cells secrete higher levels of pro-survival and immunomodulatory growth factors120. Sci. Attachment of human iPSCs was successful in assemblies functionalized with collagen I and laminin-521 but not with fibronectin. Qian, X. Y., Nguyen, H. N., Jacob, F., Song, H. J. Alginate has a number of advantages that make it attractive as a material for further study; it is inexpensive, relatively easy to modify and functionalize, biocompatible, and has been used in a wide range of biological and materials applications106,107,108. Lee, J. S. et al. & Ahn, G. Alginate-based nanomaterials: fabrication techniques, properties, and applications. Stem Cell Res. We will discuss three main alternative media: (a) decellularized extracellular matrix and other derived proteins, (b) synthetic hydrogels, which generally incorporate cell-adhesive domains or proteolytic degradation sites, and (c) gel-forming recombinant peptides. However, these studies observed the effects of pore size on mature cells rather than organoid development from multipotent cells. Hepatology 53, 604617 (2011). Rev. Light-sensitive polyvinyl alcohol matrices have recently been developed for cell culture; these matrices allow for control over the spacing of biochemical cues and the material environment134. In a follow-up study, murine cardiomyocyte differentiation (measured by -myosin expression, cell contractility, and metabolic activity) was found to be dependent on the stoichiometric ratio of THPC to protein, which tuned the stiffness of the hydrogel. Clin. Dahl-Jensen, S. & Grapin-Botton, A. Cell-binding cues from collagen, fibronectin, or laminin have frequently been added to synthetic hydrogels to allow for organoid growth and differentiation. Lin and colleagues reported that liver tissue decellularization supported growth and maintenance of rat hepatocytes; however, this method relied on mechanical disruption of resected tissue, which resulted in the loss of organ architecture and vascular networks73. 76, 7798 (2005). Front. Batchelder, C. A., Martinez, M. L. & Tarantal, A. F. Natural scaffolds for renal differentiation of human embryonic stem cells for kidney tissue engineering. A. For these reasons, organoids are used to study organ development2 and model various diseases such as cancers3, neural disorders4, and autism5; they are also used as pharmaceutical testing platforms6, model systems for CRISPR-CAS9-mediated treatment of genetic diseases7, and replacement organs for transplantation8,9. A 3D Alzheimers disease culture model and the induction of P21-activated kinase mediated sensing in iPSC derived neurons. Eng. https://doi.org/10.1038/s41580-020-0259-3 (2020). 14, 523531 (2015). Stem cells and/or organ progenitors from normal or diseased tissue can be mixed with Matrigel matrix or Collagen to create mini-organs of the kidney, thyroid, liver, brain, lung, intestine, prostate, and pancreas. Establishment of a human iPSC- and nanofiber-based microphysiological blood-brain barrier system. The human pancreas presents a challenge in decellularization as it has a higher lipid content than animal models. Standard cell culture procedures including sterile handling in Class II biosafety cabinets and incubation in humidified incubators at 37C and 5% CO 2 are required for maintenance of hPSC lines. 1500Pa, as indicated by higher expression of -2 tubulin and microtubule-associated protein 2. Biomaterials 233, 119673 (2020). PubMed We also thank Prof. David A. Tirrell for helpful discussions and editing assistance. https://doi.org/10.3791/54331 (2016). Chung and colleagues generated a hydrogel using an elastin-like polypeptide matrix containing a fibronectin-derived RGD cell-binding domain with tetrakis(hydroxymethyl)phosphonium chloride (THPC) as an amine-reactive crosslinker; this material transiently inhibited contractility of murine ESC-derived cardiomyocytes and enhanced survival of dorsal root ganglia cells from chick embryos189. Stem Cells Transl. Dye and coworkers found that human lung organoids transplanted in mice could merge together to form airway structures. Development 144, 968975 (2017). Gjorevski, N. et al. We will review undefined matrices, focusing on ECM derived from decellularized tissues and collagen, and defined matrices, including synthetic polymer hydrogels and engineered ECM proteins (Fig. PubMed Elasticity-based development of functionally enhanced multicellular 3D liver encapsulated in hybrid hydrogel. Figure 7. Wang, Y. et al. Concordance of the toxicity of pharmaceuticals in humans and in animals. 11, 16621674 (2010). https://doi.org/10.1016/j.semcdb.2021.03.011, https://doi.org/10.1038/s41580-020-0259-3, https://www.corning.com/catalog/cls/documents/application-notes/CLS-AC-AN-449.pdf, https://doi.org/10.18632/oncotarget.10945, http://creativecommons.org/licenses/by/4.0/. Table2 summarizes studies that discuss the effects of elastic modulus on the organoid formation of various tissues. Nonadhesive alginate hydrogels support growth of pluripotent stem cell-derived intestinal organoids. Decellularized ECM has been prepared from various pancreatic cell sources, such as adult human pancreas81 and porcine pancreas82. USA 116, 74837492 (2019). Intriguingly, when PEG was crosslinked with dithiothreitol to inhibit matrix degradation, organoid viability at seven days was poor as measured by live-dead staining. Tech. 1. & Gelain, F. Branched peptides integrate into self-assembled nanostructures and enhance biomechanics of peptidic hydrogels. 5, 289298 (2018). In situ cell manipulation through enzymatic hydrogel photopatterning. Biomacromolecules 4, 713722 (2003). 3D niche microarrays for systems-level analyses of cell fate. Mark T. Kozlowski. Biomacromolecules 13, 39123916 (2012). Protein-based hydrogels can be outfitted with a broad range of chemical functionalities by introducing noncanonical amino acids208,209; they can also be readily tailored to a wide variety of biomedical contexts210,211 and made thermally responsive212,213. Front. 5, 159160 (2018). Towards organoid culture without Matrigel - PMC - National Center for 4 integrin expressed by intestinal organoids is distributed only at the basal surface94, while 1 integrin is required for proper apical-basal polarization95,96,97. What are Matrigel domes? Mater. and H.T.K. A similar process was used by Nair and colleagues using mechanical agitation to keep human cells suspended in culture, resulting in functional beta-like clusters221. 4, 534539 (2015). Coat inserts with 3% BSA in PBS (approx. 5. Lee, S. S. et al. Co, J. Y. et al. In another follow-up study, a very soft matrix with an elastic modulus of 180Pa promoted intestinal organoid-forming efficiencies comparable to those observed in collagen I-based matrices. A. A., Lutolf, M. P. & Martinez-Arias, A. Nat. Chimia 71, 342348 (2017). Cornings new Matrigel matrix for organoid culture is an optimized version of this trusted product, formulated to support organoid growth and differentiation. The image is from ref. After 2 weeks, endocrine-acinar organoids were observed, demonstrating that aECM-lam was capable of inducing differentiation of ductal progenitor cells into endocrine and acinar lineages192. Brain organoids: an ensemble of bioassays to investigate human neurodevelopment and disease. Patra, B. et al. Reliability of human cortical organoid generation. 6, 13581370 (2018). Hynes, R. O. Toxicol. Developing defined substrates for stem cell culture and differentiation. 38, 220 (2010). Wu, S. H., Xu, R. J., Duan, B. 11. PDF Corning Matrigel Matrix for Organoid Culture, Phenol Red-free, 10 mL Driehuis, E. et al. After successful engraftment, HIOs generated from 4-arm PEG maleimide promoted the healing of mucosal wounds in a mouse colon injury model. A related difficulty is that not all decellularization protocols are equally effective at removing cells or other immunogenic species, which can cause varying host immune responses and failure of implants in clinical trials122. This section describes the steps required to efficiently maintain, culture, passage, and expand CRC organoids for establishing CRC organoid lines. The authors found that stiffer gels drove pancreatic progenitor cells to aggregate, leading to increased differentiation and maturation into beta-like cells; this may have been mediated through paracrine signaling enhanced by cellular proximity. The Tokyo Medical and Dental University (TMDU) method uses collagen I for intestinal enteroid culture89. The authors hypothesized that cells create their own niche within the alginate hydrogel by secreting basement membrane proteins and forming mesenchyme, allowing cellular survival and differentiation into HIOs103. Cell Press: STAR Protocols Methods Mol. Cell 155, 357368 (2013). Using aECM-lam, other morphologically-distinct organoids were formed from murine postnatal pancreas194 and sorted adult ductal progenitor cells195. In vitro expansion of single Lgr5(+) liver stem cells induced by Wnt-driven regeneration. Alginate formulations: current developments in the race for hydrogel-based cardiac regeneration. PubMedGoogle Scholar. Biopolymers 94, 6077 (2010). Biomaterials 234, 119757 (2020). Cell 161, 933945 (2015). Learn More. (d) Organoids grown in aECM-lam are capable of secreting insulin in vitro when challenged by high concentrations of d-glucose or a combination of d-glucose and cAMP activator theophylline. Step-by-step protocol for on-top MDCK 3D culture and embedded MDCK 3D culture. For example, emphysematous or fibrotic lung tissue has hardened and undergone alterations in its architecture. Mosiewicz, K. A. et al. The authors declare no competing interests. 5, 15 (2014). Biomater. Curr. Download: Protocol for Corning Matrigel Basement Membrane Matrix for 3D Culture In Vitro. 2.1 HIO Culture. Shimizu, T., Yamagata, K. & Osafune, K. Kidney organoids: research in developmental biology and emerging applications. Enhanced invasion of metastatic cancer cells via extracellular matrix interface. Corning Matrigel matrix is the most published hydrogel for organoid research due to its close resemblance to an in vivo environment, providing necessary growth factors, proteins, and the required matrix architecture. Am. Schwartz and colleagues used a PEG-based gel functionalized with pendant RGD cell-binding domains and crosslinked by MMP-degradable peptides to generate neural organoids169. Matrigel matrix for organoids is an ideal ECM for culturing 3D organoid models. Pancreatic cancer organoids recapitulate disease and allow personalized drug screening. Cell Develop. This study represents a new direction towards the generation of organoids, where the shape of the culture vessel might be tuned in order to change cellular phenotype, growth, and differentiation. PLoS ONE 10, 18 (2015). Annu. Cell Stem Cell 22, 45445 (2018). Hydrogel-based materials could be engineered in order to locally provide specific biochemical and mechanical signals able to trigger lineage commitment, spatial . A 3D culture in RADA-16 resulted in activation of a p21-activated kinase in response to amyloid- oligomers. Nature 499, 48148 (2013). From background product information to method instructions, discover guidelines for using Matrigel matrix for organoid culture. Polym. Wilkinson, D. C. et al. Nat. Innovative encapsulation platform based on pancreatic extracellular matrix achieve substantial insulin delivery. Wagner, D. E. et al. Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. Tabata, Y.
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