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人脐带间充质干细胞在与睾丸组织细胞共培养的作用下向类生殖细胞分化。

Differentiation of human umbilical cord mesenchymal stem cell into germ-like cell under effect of co-culture with testicular cell tissue.

机构信息

Department of Animal Physiology, Shahid Beheshti University, Tehran, Iran.

Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.

出版信息

Anat Histol Embryol. 2020 May;49(3):359-364. doi: 10.1111/ahe.12537. Epub 2020 Feb 7.

Abstract

Supplements produced by mouse testicular cells (mTCs) and the interaction between cells can increase the differentiation rate of human umbilical cord mesenchymal stem cells (hUCMSCs) into the germ-like cells. We studied the differentiation rate of hUCMSCs into the germ-like cells under effect of mTCs co-culturing. Isolated hUCMSCs from postpartum human umbilical cords were cultured. Then, the expression of mesenchymal (CD73, CD90 and CD105) and haematopoietic (CD34 and CD45) markers of hUCMSCs were confirmed by flow cytometry. Then, the hUCMSCs were cultured in four distinct groups: (a) control, (b) co-culture until D0, (c) co-culture until D5 and (d) co-culture until D10, in order to differentiate into the germ-like cells. After 10 days, the expression of OCT4, VASA, Fragilis and SYCP3 genes were examined by Real-Time qPCR. The flow cytometry indicated a high expression of mesenchymal markers and a low expression of haematopoietic markers (CD73:98.6%, CD90: 99.1%, CD105: 99.5%, CD34: 4.22% and CD45: 2.54%). The expression of OCT4 decreased during the time while the expression of VASA, Fragilis and SYCP3 markers increased in the co-culture with testicular cells (p value <.05). Co-culture with mTCs may be used as an effective method to differentiate hUCMSCs into germ-like cells.

摘要

由小鼠睾丸细胞(mTCs)产生的补充剂和细胞之间的相互作用可以提高人脐带间充质干细胞(hUCMSCs)向类生殖细胞分化的分化率。我们研究了 mTCs 共培养对 hUCMSCs 向类生殖细胞分化的影响。从产后人脐带中分离 hUCMSCs 进行培养。然后,通过流式细胞术确认 hUCMSCs 的间充质(CD73、CD90 和 CD105)和造血(CD34 和 CD45)标志物的表达。然后,hUCMSCs 分别在四个不同的组中培养:(a)对照组;(b)共培养至 D0 天;(c)共培养至 D5 天;(d)共培养至 D10 天,以分化为类生殖细胞。10 天后,通过实时 qPCR 检测 OCT4、VASA、Fragilis 和 SYCP3 基因的表达。流式细胞术表明间充质标志物表达较高,造血标志物(CD73:98.6%、CD90:99.1%、CD105:99.5%、CD34:4.22%和 CD45:2.54%)表达较低。在共培养过程中,OCT4 的表达随着时间的推移而降低,而 VASA、Fragilis 和 SYCP3 标志物的表达在与睾丸细胞共培养时增加(p 值<.05)。mTCs 的共培养可能是将 hUCMSCs 分化为类生殖细胞的有效方法。

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