心肌细胞选择性培养基

Cardiomyocyte Selective Medium (CSM) is a complete medium designed for the purification of cardiomyocyte cells derived from human pluripotent stem cells (hPSC). Current protocols allow for the differentiation of hPSC to cardiomyocytes at a high efficiency, however, the derived cells are a mixed population containing non-cardiomyocyte cells. The non-cardiomyocyte cells may interfere with downstream analyses. Density-gradient centrifugation, genetic modification, and cell sorting based on cell surface markers or mitochondria dyes have been applied to enrich for the hPSC-derived cardiomyocytes. These methods, however, either involve complicated procedures or require special equipment, and are not ideal for therapeutic purposes. Based on the unique metabolic properties of fetal and hPSC-derived cardiomyocytes, which can use lactate as a major energy source, CSM is developed to enrich for the hPSC-derived cardiomyocytes using an efficient, easy, and non-invasive approach. CSM is a serum-free medium specially designed for the selective growth of cardiomyocytes. It significantly improves the purity of hPSC-derived cardiomyocyte cell populations in 4 - 6 days, requiring only daily medium change. While culturing in the CSM, the non-cardiomyocyte cells were gradually eliminated, whereas the cardiomyocytes remain contracting in the culture. The CSM can also be used for cardiomyocytes derived from mouse pluripotent stem cell and/or the primary cells isolated from the hearts of rat neonatal pups.

心肌细胞选择性培养基（Cardiomyocyte Selective Medium，CSM）是一种用于纯化来源于人多能干细胞（hPSC）的心肌细胞的完整培养体系。当前的分化方法虽可高效将hPSC诱导为心肌细胞，但所得细胞群体通常为混合群体，包含一定比例的非心肌细胞，可能对后续实验分析产生干扰。为提高心肌细胞纯度，研究中常采用密度梯度离心、基因修饰或基于细胞表面标志物及线粒体染料的分选方法，但这些方法往往操作复杂、依赖专用设备，且不利于临床转化应用。

CSM基于胎儿期及hPSC来源心肌细胞独特的代谢特性（可利用乳酸作为主要能量来源）而开发，提供了一种高效、简便且非侵入性的纯化策略，用于富集hPSC来源的心肌细胞。该培养基为无血清、成分明确的体系，专为心肌细胞的选择性生长而设计。在仅需每日更换培养基的条件下，可在4–6天内显著提高心肌细胞群体的纯度。在CSM培养过程中，非心肌细胞逐渐被清除，而心肌细胞则持续保持收缩活性。此外，CSM同样适用于来源于小鼠多能干细胞的心肌细胞，以及从新生大鼠心脏分离获得的原代心肌细胞培养。