Gong Seung Pyo, Lee Eun Ju, Lee Seung Tae, Kim Heebal, Lee Sang Hun, Han Ho Jae, Lim Jeong Mook
Gamete and Stem Cell Biotechnology Laboratory, Department of Agricultural Biotechnology, Seoul National University, Seoul, Korea.
Stem Cells Dev. 2008 Aug;17(4):695-712. doi: 10.1089/scd.2007.0168.
This study was conducted to improve establishing autologous embryonic stem cells (ESCs) by culture of preantral follicles and parthenogenetic activation of oocytes. First, paternal inheritance of the follicle donor was changed without altering maternal heredity by employing B6CBAF1 instead of B6D2F1 mice. A significant increase in the establishment of parthenogenetic ESCs was detected after the change, and a different gene expression pro-file was detected in the ESCs established. Among 62 stemness-related genes showing different expression level between two strains, 35 (56.5%) were lower in the rarely established ESCs (B6D2F1) than in the easily established ESCs (B6CBAF1). Several paternally expressed genes were aberrantly expressed in the rarely established ESCs. Second, the establishment of parthenogenetic ESCs in B6D2F1 was significantly improved when preantral follicles were cultured in glutathione (GSH)-containing medium. In the ESCs derived from GSH-treated follicles, 77% of the 62 genes showing the difference increased their expression. Translation of several proteins related to stemness (Wnt-1, beta-catenin, p-p44/42, and smad) was similar between the parthenogenetic ESCs established after GSH treatment and the control E14 ESCs. We concluded that change in genetic inheritance and exposure of in vitro-growing ovarian follicles to GSH contributes to improving establishment of parthenogenetic ESCs, which may help increase the feasibility of the established lines for patient-specific, stem cell therapy.
本研究旨在通过培养窦前卵泡和卵母细胞孤雌激活来改进自体胚胎干细胞(ESC)的建立。首先,通过使用B6CBAF1小鼠而非B6D2F1小鼠,在不改变母系遗传的情况下改变卵泡供体的父系遗传。改变后检测到孤雌胚胎干细胞建立的显著增加,并且在建立的胚胎干细胞中检测到不同的基因表达谱。在两个品系之间显示出不同表达水平的62个与干性相关的基因中,35个(56.5%)在难以建立的胚胎干细胞(B6D2F1)中比在容易建立的胚胎干细胞(B6CBAF1)中表达更低。几个父源表达基因在难以建立的胚胎干细胞中异常表达。其次,当在含谷胱甘肽(GSH)的培养基中培养窦前卵泡时,B6D2F1中孤雌胚胎干细胞的建立显著改善。在源自GSH处理卵泡的胚胎干细胞中,显示出差异的62个基因中有77%的表达增加。GSH处理后建立的孤雌胚胎干细胞与对照E14胚胎干细胞之间,几种与干性相关的蛋白质(Wnt-1、β-连环蛋白、p-p44/42和smad)的翻译相似。我们得出结论,遗传继承的改变以及体外生长的卵巢卵泡暴露于GSH有助于改善孤雌胚胎干细胞的建立,这可能有助于提高所建立细胞系用于患者特异性干细胞治疗的可行性。