Efficient derivation of human cardiac precursors and cardiomyocytes from pluripotent human embryonic stem cells with small molecule induction.
Efficient derivation of human cardiac precursors and cardiomyocytes from pluripotent human embryonic stem cells with small molecule induction.
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DOI：
10.3791/3274
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发表时间：
2011-11-03
期刊：
影响因子：
--
通讯作者：
Moore DA
中科院分区：
文献类型：
--
作者：
Parsons XH;Teng YD;Parsons JF;Snyder EY;Smotrich DB;Moore DA
迄今为止，无论是通过细胞移植还是心脏组织工程，缺乏合适的人类心脏细胞来源一直是人类心肌再生的主要障碍1-3。心肌细胞在出生后不久就会发生终末分化，并失去增殖能力。没有证据表明来自其他来源（如骨髓或脐带血）的干细胞/祖细胞在移植到心脏后能够产生可收缩的心肌细胞1-3。成体干细胞治疗无法满足再生或修复受损心肌的需要，无论是内源性的还是通过细胞输送的。遗传稳定的人类胚胎干细胞（hESCs）具有无限的扩增能力和不受限制的可塑性，为体外衍生大量需要修复和再生的人类体细胞提供了一个多能库4,5。由于全球范围内心血管疾病的流行和供体器官的严重短缺，人们对开发基于hesc的治疗方法作为一种替代方法有着浓厚的兴趣。然而，如何有效和可预测地将多能hESCs的广泛分化潜力引导到所需的表型一直是发育研究和临床转化的主要挑战。传统的方法依赖于多能细胞通过自发生殖层分化的多系倾向，导致低效和不可控的谱系承诺，通常伴随着表型异质性和不稳定性，因此具有很高的致瘤风险6-8（见图1A中的示意图）。此外，通常用于分离、扩增和分化hESCs的未定义的外来/动物生物补充剂和/或喂食器可能会使患者直接使用这种细胞特化移植物存在问题9-11。为了克服这些障碍，我们已经解决了维持hESCs外胚层多能性所必需和足够的定义培养系统的要素，作为临床适用hESCs的重新衍生平台，并通过小分子有效地将这些hESCs均匀地引导到临床相关谱系12（见图1B中的示意图）。在筛选了各种小分子和生长因子后，我们发现，这种明确的条件使得烟酰胺（NAM）足以诱导多能hESCs直接分化为心内胚层，并进一步发展为成心细胞，从而高效地产生人类跳动的心肌细胞（图2）。我们定义了直接从多能hESCs诱导成心细胞的条件，而不需要干预多系胚样体阶段，从而能够在发育阶段范围内有效地获得大量人类心脏细胞，用于细胞治疗。
To date, the lack of a suitable human cardiac cell source has been the major setback in regenerating the human myocardium, either by cell-based transplantation or by cardiac tissue engineering1-3. Cardiomyocytes become terminally-differentiated soon after birth and lose their ability to proliferate. There is no evidence that stem/progenitor cells derived from other sources, such as the bone marrow or the cord blood, are able to give rise to the contractile heart muscle cells following transplantation into the heart1-3. The need to regenerate or repair the damaged heart muscle has not been met by adult stem cell therapy, either endogenous or via cell delivery1-3. The genetically stable human embryonic stem cells (hESCs) have unlimited expansion ability and unrestricted plasticity, proffering a pluripotent reservoir for in vitro derivation of large supplies of human somatic cells that are restricted to the lineage in need of repair and regeneration4,5. Due to the prevalence of cardiovascular disease worldwide and acute shortage of donor organs, there is intense interest in developing hESC-based therapies as an alternative approach. However, how to channel the wide differentiation potential of pluripotent hESCs efficiently and predictably to a desired phenotype has been a major challenge for both developmental study and clinical translation. Conventional approaches rely on multi-lineage inclination of pluripotent cells through spontaneous germ layer differentiation, resulting in inefficient and uncontrollable lineage-commitment that is often followed by phenotypic heterogeneity and instability, hence, a high risk of tumorigenicity6-8 (see a schematic in Fig. 1A). In addition, undefined foreign/animal biological supplements and/or feeders that have typically been used for the isolation, expansion, and differentiation of hESCs may make direct use of such cell-specialized grafts in patients problematic9-11. To overcome these obstacles, we have resolved the elements of a defined culture system necessary and sufficient for sustaining the epiblast pluripotence of hESCs, serving as a platform for de novo derivation of clinically-suitable hESCs and effectively directing such hESCs uniformly towards clinically-relevant lineages by small molecules12 (see a schematic in Fig. 1B). After screening a variety of small molecules and growth factors, we found that such defined conditions rendered nicotinamide (NAM) sufficient to induce the specification of cardiomesoderm direct from pluripotent hESCs that further progressed to cardioblasts that generated human beating cardiomyocytes with high efficiency (Fig. 2). We defined conditions for induction of cardioblasts direct from pluripotent hESCs without an intervening multi-lineage embryoid body stage, enabling well-controlled efficient derivation of a large supply of human cardiac cells across the spectrum of developmental stages for cell-based therapeutics.
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DOI：
10.1073/pnas.0808488105
发表时间：
2008-10-28
影响因子：
11.1
作者：
Aubry, Laetitia;Bugi, Aurore;Perrier, Anselme L.
通讯作者：
Perrier, Anselme L.
影响因子：
24
作者：
Caspi, Oren;Huber, Irit;Gepstein, Lior
通讯作者：
Gepstein, Lior
影响因子：
64.8
作者：
Park, In-Hyun;Zhao, Rui;Daley, George Q.
通讯作者：
Daley, George Q.
影响因子：
56.9
作者：
Thomson, JA;Itskovitz-Eldor, J;Jones, JM
通讯作者：
Jones, JM
DOI：
10.1016/j.trre.2008.05.005
发表时间：
2009-01-01
期刊：
Transplantation reviews (Orlando, Fla.)
影响因子：
--
作者：
Zhu, Wei-Zhong;Hauch, Kip D;Laflamme, Michael A
通讯作者：
Laflamme, Michael A