Generation of Liver Disease-Specific Induced Pluripotent Stem Cells Along with Efficient Differentiation to Functional Hepatocyte-Like Cells
Generation of Liver Disease-Specific Induced Pluripotent Stem Cells Along with Efficient Differentiation to Functional Hepatocyte-Like Cells
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DOI：
10.1007/s12015-010-9189-3
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发表时间：
2010-12-01
影响因子：
4.8
通讯作者：
Baharvand, Hossein
中科院分区：
文献类型：
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作者：
Ghodsizadeh, Arefeh;Taei, Adeleh;Baharvand, Hossein
疾病特异性诱导多能干细胞(IPSCs)的出现为研究和模拟特定基因缺陷对人类肝脏发育的影响以及测试小分子或其他潜在的治疗相关肝病的方法提供了独特的机会。在这里，我们首次报道了通过逆转录病毒转导Yamanaka因子在血清和无饲养层培养条件下从患有酪氨酸血症、糖原储存疾病、进行性家族性遗传性胆汁淤积症的肝脏特异性患者和两个患有Crigler-Najjar综合征的兄弟姐妹中获得IPSCs。此外，它们被有效地分化为具有功能的肝细胞样细胞。这些IPSCs具有人类胚胎干细胞(HESCs)的特性，并成功地分化为三个类似hESC形态的谱系：传代、表面和多能标记、正常核型、DNA甲基化和分化。肝细胞系定向分化实验表明，IPSC来源的肝细胞表达肝细胞特异性标志。糖原和脂肪储存活性、白蛋白、甲胎蛋白和尿素的分泌、细胞色素P450代谢活性以及低密度脂蛋白和吲哚青绿摄取证实了它们的功能。我们的结果提供了主要证据，证明人类肝病特异性IPSCs为基于细胞的治疗、药物代谢、人类肝脏发育和肝功能衰竭疾病模型提供了一个令人兴奋的潜在场所。
The availability of disease-specific induced pluripotent stem cells (iPSCs) offers a unique opportunity for studying and modeling the effects of specific gene defects on human liver development in vitro and for testing small molecules or other potential therapies for relevant liver disorders. Here we report, for the first time, the derivation of iPSCs by the retroviral transduction of Yamanaka's factors in serum and feeder-free culture conditions from liver-specific patients with tyrosinemia, glycogen storage disease, progressive familial hereditary cholestasis, and two siblings with Crigler-Najjar syndrome. Furthermore, they were differentiated into functional hepatocyte-like cells efficiently. These iPSCs possessed properties of human embryonic stem cells (hESCs) and were successfully differentiated into three lineages that resembled hESC morphology, passaging, surface and pluripotency markers, normal karyotype, DNA methylation, and differentiation. The hepatic lineage-directed differentiation showed that the iPSC-derived hepatic cells expressed hepatocyte-specific markers. Their functionality was confirmed by glycogen and lipid storage activity, secretion of albumin, alpha-fetoprotein, and urea, CYP450 metabolic activity, as well as LDL and indocyanin green uptake. Our results provide proof of principal that human liver-disease specific iPSCs present an exciting potential venue toward cell-based therapeutics, drug metabolism, human liver development and disease models for liver failure disorders.