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利用非整合方法从烧伤患者来源的皮肤成纤维细胞中生成人诱导多能干细胞。

Generation of human-induced pluripotent stem cells from burn patient-derived skin fibroblasts using a non-integrative method.

机构信息

Burns Institute, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.

Department of Oncology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.

出版信息

Int J Mol Med. 2018 Jan;41(1):87-94. doi: 10.3892/ijmm.2017.3206. Epub 2017 Oct 23.

DOI:10.3892/ijmm.2017.3206
PMID:29115387
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5746323/
Abstract

Patient specific induced pluripotent stem cells (iPSCs) have been recognized as a possible source of cells for skin tissue engineering. They have the potential to greatly benefit patients with large areas of burned skin or skin defects. However, the integration virus-based reprogramming method is associated with a high risk of genetic mutation and mouse embryonic fibroblast feeder-cells may be a pollutant. In the present study, human skin fibroblasts (HSFs) were successfully harvested from patients with burns and patient-specific iPSCs were generated using a non-integration method with a feeder-free approach. The octamer-binding transcription factor 4 (OCT4), sex-determining region Y box 2 (SOX2) and NANOG transcription factors were delivered using Sendai virus vectors. iPSCs exhibited representative human embryonic stem cell-like morphology and proliferation characteristics. They also expressed pluripotent markers, including OCT4, NANOG, SOX2, TRA181, stage-specific embryonic antigen 4 and TRA-160, and exhibited a normal karyotype. Teratoma and embryoid body formation revealed that iPSCs were able to differentiate into cells of all three germ layers in vitro and in vivo. The results of the present study demonstrate that HSFs derived from patients with burns, may be reprogrammed into stem cells with pluripotency, which provides a basis for cell‑based skin tissue engineering in the future.

摘要

患者特异性诱导多能干细胞(iPSCs)已被认为是皮肤组织工程细胞的可能来源。它们有可能极大地造福于大面积烧伤或皮肤缺陷的患者。然而,整合病毒的重编程方法存在较高的基因突变风险,而鼠胚胎成纤维细胞饲养细胞可能是一种污染物。在本研究中,成功地从烧伤患者中分离出了人皮肤成纤维细胞(HSFs),并使用无整合的方法,在无饲养细胞的条件下生成了患者特异性 iPSCs。八聚体结合转录因子 4(OCT4)、性别决定区 Y 框 2(SOX2)和 NANOG 转录因子通过仙台病毒载体传递。iPSCs 表现出具有代表性的人类胚胎干细胞样形态和增殖特征。它们还表达多能性标记物,包括 OCT4、NANOG、SOX2、TRA181、阶段特异性胚胎抗原 4 和 TRA-160,并表现出正常的核型。畸胎瘤和类胚体形成表明 iPSCs 能够在体外和体内分化为三个胚层的细胞。本研究的结果表明,来自烧伤患者的 HSFs 可以被重编程为具有多能性的干细胞,为未来的基于细胞的皮肤组织工程提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/caf850e2468a/IJMM-41-01-0087-g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/8a80ff723ca7/IJMM-41-01-0087-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/7b21a59d1b69/IJMM-41-01-0087-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/3a80bad779c5/IJMM-41-01-0087-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/ae3d62a545ae/IJMM-41-01-0087-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/6916ad2c51bd/IJMM-41-01-0087-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/caf850e2468a/IJMM-41-01-0087-g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/8a80ff723ca7/IJMM-41-01-0087-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/7b21a59d1b69/IJMM-41-01-0087-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/3a80bad779c5/IJMM-41-01-0087-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/ae3d62a545ae/IJMM-41-01-0087-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/6916ad2c51bd/IJMM-41-01-0087-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b47/5746323/caf850e2468a/IJMM-41-01-0087-g05.jpg

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