Soong Poh Loong, Tiburcy Malte, Zimmermann Wolfram-Hubertus
Department of Pharmacology, Heart Research Center Göttingen (HRCG), University Medical Center Göttingen, Göttingen, Germany.
Curr Protoc Cell Biol. 2012 Jun;Chapter 23:Unit23.8. doi: 10.1002/0471143030.cb2308s55.
The advent of pluripotent human embryonic stem cells has created the unique opportunity for the development of a wide variety of humanized cellular tools for basic research, as well as industrial and clinical applications. It has, however, become apparent that embryonic stem cell derivatives in classical monolayer or embryoid body culture do not resemble bona fide tissues, mainly because of their limited organotypic organization and maturation in these culture formats. This shortcoming may be addressed by tissue engineering technologies aiming at the provision of a "natural" growth environment to facilitate organotypic tissue assembly. In this unit, we provide two harmonized basic protocols for (1) cardiac differentiation of human embryonic stem cells under serum-free conditions and (2) the assembly of the stem cell-derived cardiomyocytes into engineered heart muscle. This protocol can be easily adapted to bioengineer heart muscle also from other stem cell-derived cardiomyocytes, including cardiomyocytes from human-induced pluripotent stem cells.
多能性人类胚胎干细胞的出现为开发用于基础研究以及工业和临床应用的各种人源化细胞工具创造了独特的机会。然而，很明显，经典单层或胚状体培养中的胚胎干细胞衍生物并不类似于真正的组织，主要是因为它们在这些培养形式中的器官样组织和成熟有限。这个缺点可以通过组织工程技术来解决，这些技术旨在提供一个“自然”的生长环境以促进器官样组织的组装。在本单元中，我们提供了两个协调的基本方案，用于（1）在无血清条件下对人类胚胎干细胞进行心脏分化，以及（2）将干细胞衍生的心肌细胞组装成工程化心肌。该方案可以很容易地适用于从其他干细胞衍生的心肌细胞（包括来自人类诱导多能干细胞的心肌细胞）构建生物工程心肌。
