黑种草、羽衣甘蓝和月见草提取物诱导胚胎干细胞分化为原始生殖细胞样细胞。

Embryonic stem cell differentiation to primordial germ cell like cells by Nigella sativa, Brassica Oleracea and Oenothera biennis extracts.

作者信息

Najibi Reza, Kazemitabar Seyed Kamal, Kiani Ghaffar, Hasanzadeh Nasrin, Gholami Mana, Hajimazdarany Shima, Ahmadi Ali Asghar

机构信息

Department of Biotechnology and Plant Breeding, Sari Agricultural Sciences and Natural Resources University (SANRU) Sari, Iran.

Department of Biotechnology and Plant Breeding, College of Agricultural Science, Sari Agricultural Sciences and Natural Resources University (SANRU) Sari, Iran.

出版信息

Am J Stem Cells. 2022 Dec 25;11(5):79-93. eCollection 2022.

PMID:36660740

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9845841/

Abstract

OBJECTIVES

This study aimed to investigate the induction effects of methanolic extracts of Nigella sativa (NiS), Brassica Oleracea (BrO), and Oenothera biennia (Obi) on transgenic embryonic stem cells (ESCs) and to evaluate the ability of germ cells (GCs) production using these pluripotent cells.

METHODS

ESCs were amplified using a feeder layer. Embryoid bodies enzymatically dissociated to single cells and induced the extracts in gelatinized plates. Then RNA extraction and cDNA synthesis were performed. In the presence of appropriate primers, the desired genes were quantitatively evaluated by quantitative polymerase chain reaction (qPCR).

RESULTS

The copies of all genes in the control group showed a decreasing trend during the first to third weeks. Compared to the control group, the expression level of sex determining region Y-box 2 gene (Sox2) showed the highest level. All four evaluated genes increased in all Obi groups compared to the control group. There is also a slight increase in the Nanog homeobox gene (Nanog). Obi extract in different concentrations has increased the expression of the Sox2 gene. Increased expression of this gene along with octamer-binding transcription factor 4 gene (Oct4) and Nanog indicates a condition close to germ cell-like cells (GCLCs).

CONCLUSIONS

According to the results of this study, NiS can increase expression of the Oct4, Sox2, Nanog, and stimulated by retinoic acid gene 8 (STRA8) genes and so increase the hope of GCs production. Storage of cells for 21 days in the presence of the extract compared to 14 days has a negative effect on cell growth and differentiation. The effects of meiosis onset and GCs production can be expected in the presence of some herbal extracts. Optimal utilization of these extracts requires further study in the field of different extracts and fractions of each extract to more effectively and purposefully direct the differentiation of stem cells.

摘要

目的

本研究旨在探讨黑种草（NiS）、羽衣甘蓝（BrO）和月见草（Obi）甲醇提取物对转基因胚胎干细胞（ESCs）的诱导作用，并评估利用这些多能细胞产生生殖细胞（GCs）的能力。

方法

使用饲养层扩增胚胎干细胞。将胚状体酶解为单细胞，并在明胶包被的培养皿中用提取物进行诱导。然后进行RNA提取和cDNA合成。在合适引物存在的情况下，通过定量聚合酶链反应（qPCR）对所需基因进行定量评估。

结果

对照组所有基因的拷贝数在第一至第三周呈下降趋势。与对照组相比，性别决定区Y盒2基因（Sox2）的表达水平最高。与对照组相比，所有Obi组中所有四个评估基因均增加。同源盒基因Nanog也有轻微增加。不同浓度的Obi提取物增加了Sox2基因的表达。该基因与八聚体结合转录因子4基因（Oct4）和Nanog的表达增加表明接近生殖细胞样细胞（GCLCs）的状态。

结论

根据本研究结果，NiS可增加Oct4、Sox2、Nanog和视黄酸刺激基因8（STRA8）基因的表达，从而增加产生生殖细胞的希望。与14天相比，在提取物存在下将细胞储存21天对细胞生长和分化有负面影响。在某些草药提取物存在的情况下，可以预期减数分裂开始和生殖细胞产生的效果。要更有效且有目的地指导干细胞分化，需要在不同提取物及其各部分领域进一步研究这些提取物的最佳利用方法。