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  • Human cardiomyocyte progenitor cells differentiate into functional mature cardiomyocytes: an in vitro model for studying human cardiac physiology and pathophysiology.

Human cardiomyocyte progenitor cells differentiate into functional mature cardiomyocytes: an in vitro model for studying human cardiac physiology and pathophysiology.

Nature protocols (2009-02-07)
Anke M Smits, Patrick van Vliet, Corina H Metz, Tom Korfage, Joost Pg Sluijter, Pieter A Doevendans, Marie-José Goumans
摘要

To date, there is no suitable in vitro model to study human adult cardiac cell biology. Although embryonic stem cells are able to differentiate into cardiomyocytes in vitro, the efficiency of this process is very low. Other methods to differentiate progenitor cells into beating cardiomyocytes rely on coculturing with rat neonatal cardiomyocytes, making it difficult to study human cardiomyocyte differentiation and (patho)physiology. Here we have developed a method for efficient isolation and expansion of human cardiomyocyte progenitor cells (CMPCs) from cardiac surgical waste or alternatively from fetal heart tissue. Furthermore, we provide a detailed in vitro protocol for efficient differentiation of CMPCs into cardiomyocytes with great efficiency (80-90% of differentiation). Once CMPCs are rapidly dividing ( approximately 1 month after isolation), differentiation can be achieved in 3-4 weeks.

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Sigma-Aldrich
hBFGF, FGF-Basic, recombinant, expressed in E. coli, suitable for cell culture
Sigma-Aldrich
单克隆 抗-α-肌动蛋白(肌小节) 小鼠抗, clone EA-53, ascites fluid
Sigma-Aldrich
5-阿扎胞苷, ≥98% (HPLC)
Sigma-Aldrich
明胶 来源于猪皮肤, gel strength 80-120 g Bloom, Type A
Sigma-Aldrich
乙二胺四乙酸 二钠盐 二水合物, ACS reagent, 99.0-101.0%