Efficient differentiation of human ES and iPS cells into cardiomyocytes on biomaterials under xeno-free conditions
Efficient differentiation of human ES and iPS cells into cardiomyocytes on biomaterials under xeno-free conditions
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DOI：
10.1039/c9bm00817a
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发表时间：
2019-12-01
影响因子：
6.6
通讯作者：
Higuchi, Akon
中科院分区：
文献类型：
--
作者：
Sung, Tzu-Cheng;Liu, Cheng-Hui;Higuchi, Akon
目前用于将hPSC（人多能干细胞）分化成心肌细胞的无异种和化学限定的方法是无效的，并且有时不可再现。因此，有必要开发可靠和有效的方法将hPSC分化为心肌细胞，用于未来与药物发现，心脏毒性筛选和疾病建模相关的心血管研究。我们评估了先前报道的两种代表性的分化方法，并且我们进一步开发了在无异种、化学限定的条件下将hPSC分化为心肌细胞的原始、更有效的方法。开发的方案成功地将hPSC分化为心肌细胞，其中约90-97%表达心脏标志物cTnT，其跳动速度和肌节长度与健康成人心脏相似。还使用细胞外基质模拟材料包被的培养皿评估了用于hPSC心脏分化的最佳细胞培养生物材料。根据对hPSC衍生的心肌细胞的观察，发现Synthemax II包被的和Laminin-521包被的培养皿是用于hPSC的心脏分化的最有效和高效的生物材料，所述hPSC衍生的心肌细胞具有高存活率、高搏动集落数、与健康成人心脏相似的搏动频率，更高的纯度水平（高cTnT表达）和更长的肌节长度，与健康成人心脏相似。
Current xeno-free and chemically defined methods for the differentiation of hPSCs (human pluripotent stem cells) into cardiomyocytes are not efficient and are sometimes not reproducible. Therefore, it is necessary to develop reliable and efficient methods for the differentiation of hPSCs into cardiomyocytes for future use in cardiovascular research related to drug discovery, cardiotoxicity screening, and disease modeling. We evaluated two representative differentiation methods that were reported previously, and we further developed original, more efficient methods for the differentiation of hPSCs into cardiomyocytes under xeno-free, chemically defined conditions. The developed protocol successively differentiated hPSCs into cardiomyocytes, approximately 90-97% of which expressed the cardiac marker cTnT, with beating speeds and sarcomere lengths that were similar to those of a healthy adult human heart. The optimal cell culture biomaterials for the cardiac differentiation of hPSCs were also evaluated using extracellular matrix-mimetic material-coated dishes. Synthemax II-coated and Laminin-521-coated dishes were found to be the most effective and efficient biomaterials for the cardiac differentiation of hPSCs according to the observation of hPSC-derived cardiomyocytes with high survival ratios, high beating colony numbers, a similar beating frequency to that of a healthy adult human heart, high purity levels (high cTnT expression) and longer sarcomere lengths similar to those of a healthy adult human heart.