山羊胚胎心脏成纤维细胞制备无转基因诱导多能干细胞。

IF 1.1 Q4 CELL & TISSUE ENGINEERING Journal of Stem Cells & Regenerative Medicine Pub Date : 2020-12-11 eCollection Date: 2020-01-01 DOI:10.46582/jsrm.1602007
Mira Hanna, Raja Ghazanfar Ali Sahito, Moshira Rateb, Allah Bux Kachiwal, Hanan A Seddiek, Bachal Bhutto, Jürgen Hescheler
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引用次数: 2

摘要

诱导多能干细胞(iPSCs)在治疗性再生医学中具有巨大的潜力。本研究的目的是从山羊胚胎心脏组织来源的成纤维细胞中获得诱导多能干细胞。山羊胚胎心脏组织分离成纤维细胞α -平滑肌肌动蛋白、波形蛋白和盘状蛋白结构域受体阳性。从这些细胞中,我们使用piggyBac转座子/转座酶使用五种转录因子(Oct4, Sox2, Klf, Myc和Lin 28)生成无转基因iPSCs。生成的iPSCs为SSEA1、SSEA4和Oct4阳性。用bFGF替代20%血清IMDM对新饲者进行培养。在不含bFGF的20% FBS-IMDM培养基中培养,可形成囊状致密的胚状体,并分化为外胚层样细胞和中胚层样细胞。在这种方法的框架下产生的iPSCs是在没有使用整合病毒的情况下产生的,并且在该过程结束时去除重编程转基因。虽然所使用的方法存在局限性,但获得了重编程的实质性迹象。
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Generation of transgene-free induced pluripotent stem cells from cardiac fibroblasts of goat embryos.

Induced pluripotent stem cells (iPSCs) hold a great potential for therapeutic regenerative medicine. The aim of this study was to generate induced pluripotent stem cells from goat embryonic cardiac tissue derived fibroblasts. The isolated cardiac fibroblasts from the cardiac tissue of goat embryos were positive for alfa smooth muscle actin, vimentin and discoidin domain receptor2. From these cells, we generated transgene free iPSCs using piggyBac transposons / transposase using five transcription factors (Oct4, Sox2, Klf, Myc and Lin 28). The generated iPSCs were SSEA1, SSEA4 and Oct4 positive. They were cultured on neofeeders using 20% Serum replacement - IMDM with bFGF. They could form cystic and compact embryoid bodies that showed differentiated ectodermal and mesodermal like cells when cultured using 20% FBS-IMDM without bFGF. The iPSCs, generated in the frame of this approach were produced without the use of integrating virus and the reprogramming transgenes were removed at the end of the process. Though there were limitations in the approach used, a substantial sign of reprogramming was obtained.

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来源期刊
CiteScore
3.40
自引率
0.00%
发文量
5
审稿时长
14 weeks
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