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

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

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

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

Authors

Reza Najibi¹, Seyed Kamal Kazemitabar¹, Ghaffar Kiani², Nasrin Hasanzadeh³, Mana Gholami⁴, Shima Hajimazdarany⁵, Ali Asghar Ahmadi³

Affiliations

¹ Department of Biotechnology and Plant Breeding, Sari Agricultural Sciences and Natural Resources University (SANRU) Sari, Iran.
² Department of Biotechnology and Plant Breeding, College of Agricultural Science, Sari Agricultural Sciences and Natural Resources University (SANRU) Sari, Iran.
³ North Research Center, Pasteur Institute of Iran Amol, Iran.
⁴ Department of Biology, Faculty of Science, Science and Research Branch, Islamic Azad University Tehran, Iran.
⁵ Department of Biology, Faculty of Science, Babol Branch, Islamic Azad University Babol, Iran.

PMID: 36660740
PMCID: 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.

Keywords: Brassica Oleracea; Mouse embryonic stem cells; Nigella sativa; Oenothera biennia; differentiation; germ cells; herbal medicine.