Lecture+14_Beaudin_2014

Tissue engineering requirements for success l

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Unformatted text preview: idence for “cardiac stem cells” in the mature heart Generating cardiomyocytes/cardiac progenitors from human stem cells: Current limitations l༆  Many questions remain about heart development l༆  Developmental pathways used to define markers/ growth factors/ etc for ES differentiation l༆  Lineage progression of diverse cell types l༆  Mechanisms underlying diversification of cardiac lineages l༆  Developmental potential of diverse cardiac progenitors Heart development: An overview The heart includes many distinct mature cell types, including pacemaker, valvular, arterial and venous smooth muscle, endothelial cells, and discrete atrial and ventricular cardiomyocytes Diversity of cardiac progenitors Generating CM/CPCs from human stem cells: Current limitations l༆  Many questions remain about heart development l༆  Developmental pathways used to define markers/growth factors/ etc for ES differentiation l༆  Lineage progression of diverse cell types l༆  Lack of cell- surface markers to identify cardiac progenitors and mature cells l༆  Unlike some other organs, cardiac cells are stationary and don’t interact other cell types l༆  Methods needed to generate CMs or CPCs with high efficiency and purity Approaches to generating and purifing CM from ES cells l༆  Directed differentiation l༆  Genetic manipulation l༆  Physical separation Generating and purifying CMs/ CPCs: Directed Differentiation •  Yang et al, 2008 Generating and purifying CMs/ CPCs: Directed Differentiation •  Different aim – developmental underpinnings •  Drawbacks: •  Undesired effects of growth factors •  Efficiency and PURITY Directed differentiation: Identifying markers to purify CM/CPC l༆  Use developmental biology to try to identify new markers Cardiac expression of Flk1 in E8.5 embryos Cardiac expression of PDGFRα in E8 embryos FHF - /- Flk1 +/+ PDGFRα cTnt+ cells grown on cardiac fibroblast feeders Generating and purifying CMs: Genetic manipulations l༆  Addition of genetic markers (Nkx2.5, mhc, antibiotic selection) Elliot et al, Nat Methods, 2011 l༆  Advantages: l༆  Identification, purificatio...
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