Generation of human iPSCs from human peripheral blood mononuclear cells using non-integrative Sendai virus in chemically defined conditions.
Generation of human iPSCs from human peripheral blood mononuclear cells using non-integrative Sendai virus in chemically defined conditions.
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DOI：
10.1007/978-1-62703-511-8_7
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Wu JC
中科院分区：
文献类型：
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作者：
Churko JM;Burridge PW;Wu JC
人类诱导多能干细胞（hiPSC）由于其分化为多种细胞类型的能力而受到极大的关注，这些细胞类型表现出患者的原始表型。hiPSC的使用对于心脏生物学的研究特别有价值，因为人类心肌细胞难以分离和培养并且具有有限的增殖潜力。通过从患有心脏病的患者中获得iPSC，并随后将这些hiPSC分化为心肌细胞，可以在体外研究心脏生物学并建立心脏疾病模型。虽然有许多不同的方法用于衍生hiPSC，但这些hiPSC的临床使用将需要通过不涉及原始基因组修饰（非整合）或掺入可能含有异种试剂的异种衍生产物（如牛血清白蛋白）的方法衍生。理想情况下，该推导将在化学定义的条件下进行，以防止批次间变异性并提高重现性。此外，从细胞类型如成纤维细胞衍生需要延长培养（4-6周），大大增加了从活检进展到hiPSC所需的时间。在本文中，我们概述了培养外周血单核细胞（PBMC）并使用非整合仙台病毒将PBMC重编程为hiPSC的方法。
Human induced pluripotent stem cells (hiPSCs) have received enormous attention because of their ability to differentiate into multiple cell types that demonstrate the patient’s original phenotype. The use of hiPSCs is particularly valuable to the study of cardiac biology, as human cardiomyocytes are difficult to isolate and culture and have a limited proliferative potential. By deriving iPSCs from patients with heart disease, and subsequently differentiating these hiPSCs to cardiomyocytes, it is feasible to study cardiac biology in vitro and model cardiac diseases. While there are many different methods for deriving hiPSCs, clinical use of these hiPSCs will require derivation by methods that do not involve modification of the original genome (non-integrative) or incorporate xeno-derived products (such as bovine serum albumin) which may contain xeno-agents. Ideally, this derivation would be carried out under chemically-defined conditions to prevent lot-to-lot variability and enhance reproducibility. Additionally, derivation from cell types such as fibroblasts requires extended culture (4–6 weeks), greatly increasing the time required to progress from biopsy to hiPSC. Herein, we outline a method of culturing peripheral blood mononuclear cells (PBMCs) and reprogramming PBMCs into hiPSCs using a non-integrative Sendai virus.