The effects of sertoli cell condition medium and retinoic acid on cell viability and apoptotic morphology of bone marrow mesenchymal stem cells

سال انتشار: 1396
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 350

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شناسه ملی سند علمی:

ISERB03_064

تاریخ نمایه سازی: 11 خرداد 1397

چکیده مقاله:

Background: Differentiation of germ cells from other cell source could have great application for treatment of infertility. In this study, the effects of retinoic acid (as differentiation inducer) and sertoli cell condition medium were evaluated on viability and morphology of bone marrow mesenchymal stem cells.Methods: MSCs were extracted from femur and tibia bone marrow by aspiration method. The samples were transferred directly into 75-cm3 flasks and suspended in low-glucose DMEM containing 10% fetal calf serum (FBS) and 5% (v/v) CO2 at 37•C. The medium was changed every three days. After 70-80 % confluency in culture system, the cells were isolated by trypsin-EDTA and counted with trypan blue, then adjusted to desired densities (2× 10 5) into each well of a 6-well multidish in three groups: 1) in the presence of 10 -6 M retinoic acid 2) in the presence of sertoli cell conditioned medium 3) in the presence of retinoic acid + sertoli cell conditioned medium for 7 days. In the end of culture cell viability was evaluated by MTT assay test. Morphology of the apoptotic cells in culture system was assayed by etidium bromide/acridin orange (EB/AO) staining and DAPI test.Result: The results revealed that rate of apoptotic cells were significantly increased in RA only treated group on days 10 and 15 of culture (respectively 21/15 ± 1/7 and 23/25 ± 6/15) (PConclusion: With considering to differentiation effects of RA, by adding condition medium to RA only treated group, low rate of apoptotic cells are observed in culture system that it is suitable for cell trophy.

نویسندگان

Maryam Salem

University of Mohaghegh Adabili, Ardabil, Iran

Tooba Mirzapour

Faculty of Science, University of Mohaghegh Ardabili, Ardabil, Iran

Abolfazl Bayrami

Ardabil University of Medical Sciences, Ardabil, Iran

Mohsen Sagha

Ardabil University of Medical Sciences, Ardabil, Iran