Ghatreh Keihan, Eliyasi Masoumeh, Alaei Shahla, Saki Ghasem
Clinical Biochemistry Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.
Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.
Andrologia. 2021 Feb;53(1):e13892. doi: 10.1111/and.13892. Epub 2020 Nov 9.
This study aimed to culture the adipose tissue-derived mesenchymal stem cells (AT-MSCs) with and without leukaemia inhibitory factor (LIF), glial cell line-derived neurotrophic factor (GDNF), epidermal growth factor (EGF) and retinoic acid (RA), and investigate their impact on the differentiation of these cells into germ cells. MSCs were separated from adipose tissue of mice, and the nature of these cells is confirmed by flow cytometry. The cells were cultured in different conditions, including MSCs grown in the presence of the growth factors, MSCs without the growth factors, MSCs cultured with combined growth factors and RA, and MSCs cultured with RA. After 2 weeks, the gene expression of c-Kit, Gcnf, Mvh and Scp3 and the protein expression of c-Kit and Gcnf were assessed by real-time PCR and Western blot, respectively. Scp3 was overexpressed in the groups supplemented with RA (p < .01). The expression of c-Kit and Mvh in the growth factor-supplemented groups was increased (p < .01). Western blot analysis confirmed the real-time PCR results. The use of the growth factors for the long-term culture of stem cells can be beneficial. However, to promote germ cell differentiation, the growth factors might be used by other meiosis inducer factors, such as RA.
本研究旨在培养添加和不添加白血病抑制因子(LIF)、胶质细胞源性神经营养因子(GDNF)、表皮生长因子(EGF)和视黄酸(RA)的脂肪组织来源间充质干细胞(AT-MSCs),并研究它们对这些细胞向生殖细胞分化的影响。从小鼠脂肪组织中分离出间充质干细胞,并通过流式细胞术确认这些细胞的性质。将细胞在不同条件下培养,包括在生长因子存在下生长的间充质干细胞、无生长因子的间充质干细胞、与联合生长因子和RA一起培养的间充质干细胞以及与RA一起培养的间充质干细胞。2周后,分别通过实时PCR和蛋白质印迹法评估c-Kit、Gcnf、Mvh和Scp3的基因表达以及c-Kit和Gcnf的蛋白质表达。在补充RA的组中Scp3过表达(p <.01)。在补充生长因子的组中c-Kit和Mvh的表达增加(p <.01)。蛋白质印迹分析证实了实时PCR结果。使用生长因子进行干细胞的长期培养可能是有益的。然而,为了促进生殖细胞分化,生长因子可能需要与其他减数分裂诱导因子如RA一起使用。