Saeed Azandeh, Mahin Taheri Moghadam, Mohammad Rashno, Mahvash Zargar, Pardis Abed Zadeh
JBRA Assist. Reprod. 2022; 26 (2):288-298
Received March 01, 2021
Accepted September 15, 2021
Abstract
Human follicular fluid (FF) contain different cell populations including mesenchymal stem cells. Studies tried to improve their differentiation to oocyte and use for infertility goals. Using an antioxidant can improve the quality of these cells. In present study the effects of different doses of melatonin on FF derived cells and grew to oocyte-like cells (OLC) investigated for 2 weeks. Cell viability (MTT assay), flow cytometry and ICC staining utilized to evaluate CD105 and CD34 expression, colony forming unit assay (CFU-F) capability, qRT-PCR to investigate ZP1, ZP2, ZP3, GDF9, and SCP3 expression. Hormone measurement for assessment of AMH, Estradiol and Progesterone in supernatant were conducted. Morphological characteristics of fibroblast-like cells change to a round shape specifically in 10 -7 M treated group after 2 weeks. There was no difference between control and treatment groups for MTT and CFU assays. ICC staining were positive for CD105 marker and negative for CD34 hematopoietic stem cell marker. qRT-PCR results indicated that ZP1, ZP2, GDF9, and SCP3 expression increased in 10 -7 M group in week 2; while ZP3 decreased in this group. Progesterone and AMH were detected in differentiation medium. It is declared the proper dose of melatonin can improve OLCs formation in vitro.
