Hosseinzadeh Shirzeily Maryam, Pasbakhsh Parichehr, Amidi Fardin, Mehrannia Kobra, Sobhani Aligholi
Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Iran J Reprod Med. 2013 Dec;11(12):965-76.
Recent publications about differentiation of stem cells to germ cells have motivated researchers to make new approaches to infertility. In vitro production of germ cells improves understanding differentiation process of male and female germ cells. Due to the problem of using embryonic stem cells (ESC), it's necessary the mentioned cells be replaced with some adult multi-potent stem cells in laboratories.
The aim of this study was to obtain germ cells from appropriate source beyond ESC and compare differential potentials of adipocytes derived stem cells (ADMSCs) with bone marrow derived stem cells (BMMSCs).
To find multi-potential entity, after providing purified ADMSCs and BMMSCs, differentiation to osteoblast and adipocyte was confirmed by using appropriate culture medium. To confirm mesenchymal lineage production superficial markers (expression of CD90 and CD44 and non-expression of CD45 and CD31) were investigated by flowcytometry. Then the cells were differentiated to germ cells in inductive medium containing retinoic acid for 7days. To evaluate germ cells characteristic markers [Dazl (Deleted in azoospermia-like), Mvh (Mouse vasa homolog gene), Stra8 (Stimulated by retinoic acid) and Scp3 (Synaptonemal complex protein 3)] flowcytometry, imunoflorescence and real time PCR were used.
Both types of cells were able to differentiate into osteoblast and adipocyte cells and presentation of stem cell superficial markers (CD90, CD44) and absence of endothelial and blood cell markers (CD31, CD45) were confirmative The flowcytometry, imunoflorescence and real time PCR results showed remarkable expression of germ cells characteristic markers (Mvh, Dazl, Stra8, and Scp3).
It was found that although ADMSCs were attained easier and also cultured and differentiated rapidly, germ cell markers were expressed in BMMSCs significantly more than ADMSCs. This article extracted from M.Sc. thesis. (Maryam Hosseinzadeh Shirzeily).
近期有关干细胞向生殖细胞分化的出版物促使研究人员探索治疗不孕症的新方法。体外生殖细胞的产生有助于深入了解雄性和雌性生殖细胞的分化过程。由于使用胚胎干细胞(ESC)存在问题,因此实验室有必要用一些成体多能干细胞来替代这些细胞。
本研究旨在从ESC以外的合适来源获取生殖细胞,并比较脂肪来源干细胞(ADMSC)与骨髓来源干细胞(BMMSC)的分化潜能。
为寻找多能细胞,在获取纯化的ADMSC和BMMSC后,使用合适的培养基确认其向成骨细胞和脂肪细胞的分化。通过流式细胞术研究间充质谱系产生的表面标志物(CD90和CD44表达以及CD45和CD31不表达)以确认。然后将细胞在含有视黄酸的诱导培养基中分化7天以形成生殖细胞。为评估生殖细胞特征性标志物[无精子症样缺失基因(Dazl)、小鼠维甲酸同源基因(Mvh)、视黄酸刺激基因8(Stra8)和联会复合体蛋白3(Scp3)],采用了流式细胞术、免疫荧光和实时PCR技术。
两种类型的细胞都能够分化为成骨细胞和脂肪细胞,并且干细胞表面标志物(CD90、CD44)的呈现以及内皮细胞和血细胞标志物(CD31、CD45)的缺失得到了证实。流式细胞术、免疫荧光和实时PCR结果显示生殖细胞特征性标志物(Mvh、Dazl、Stra8和Scp3)有显著表达。
研究发现,尽管ADMSC更容易获得,并且培养和分化迅速,但BMMSC中生殖细胞标志物的表达明显高于ADMSC。本文摘自硕士学位论文。(玛丽安·侯赛因扎德·希尔泽利)