![]() co-culture with stromal cells or feeder cells, and on specific culture substrates:.Proper differentiation of the cell type of interest is verified by analyzing cell type specific markers, gene expression profile, and functional assays. A limited number of signaling factors such as growth factors or small molecules, controlling cell differentiation, is applied sequentially or in a combinatorial manner, at varying dosage and exposure time. For this purpose, pluripotent stem cells (PSCs) are cultured in controlled conditions involving specific substrate or extracellular matrices promoting cell adhesion and differentiation, and define culture media compositions. Directed differentiation consists in mimicking developmental (embryo's development) decisions in vitro using the stem cells as source material. Methods Ĭell differentiation involves a transition from a proliferative mode toward differentiation mode. Fibroblasts or other differentiated cell types have been used for direct reprogramming strategies. Historically, embryonic carcinoma (EC) cells have also been used. Since the discovery of embryonic stem (ES) cells (1981) and induced pluripotent stem (iPS) cells (2006), source material is potentially unlimited. It is thus possible to direct cell fate by controlling cell decisions through extracellular signaling, mimicking developmental signals.ĭirected differentiation is primarily applied to pluripotent stem cells (PSCs) of mammalian origin, in particular mouse and human cells for biomedical research applications. ![]() Cell differentiation and tissue organogenesis involve a limited set of developmental signaling pathways. Developmental biology and embryology provides the basic knowledge of the cell types' differentiation through mutation analysis, lineage tracing, embryo micro-manipulation and gene expression studies. The differentiation process can be modeled as sequence of binary decisions based on probabilistic or stochastic models. Conceptual frame ĭuring differentiation, pluripotent cells make a number of developmental decisions to generate first the three germ layers ( ectoderm, mesoderm and endoderm) of the embryo and intermediate progenitors, followed by subsequent decisions or check points, giving rise to all the body's mature tissues. Efficient directed differentiation requires a detailed understanding of the lineage and cell fate decision, often provided by developmental biology. Stem cells are by definition pluripotent, able to differentiate into several cell types such as neurons, cardiomyocytes, hepatocytes, etc. It is essentially harnessing the potential of stem cells by constraining their differentiation in vitro toward a specific cell type or tissue of interest. Directed differentiation is a bioengineering methodology at the interface of stem cell biology, developmental biology and tissue engineering. ![]()
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