Effects of Epidermal Growth Factor, Glial Cell Line-Derived Neurotrophic and Leukemia Inhibitory Factor on the Proliferation and Differentiation Potential of Adipose Tissue-Derived Mesenchymal Stem Cells

Abstract

Background: There is a great deal of interest in using adipose tissue-derived mesenchymal stem cells (AT-MSCs) for clinical applications. However, the important limitations of clinical application of stem cells are the small number of cells and their differentiation into undesirable lineage in vitro. To overcome this problem, various growth factors are studied extensively. Objectives: The current study aimed at using 3 different doses of epidermal growth factor (EGF), glial cell line-derived neurotrophic (GDNF), and leukemia inhibitory factor (LIF) to culture AT-MSCs and evaluating their effects on proliferation, viability, differentiation potential, and maintenance of the stemness state of cells. Methods: The current experimental study was conducted on 8 -10 male NMRI (Naval medical research institute) mice provided from research center and experimental animal house of Jundishapur University of Ahvaz, Iran, from September 2016 to April 2017. AT-MSCs were isolated from mice adipose tissue. The cells were cultured with three different doses of EGF, LIF, and GDNF. The morphology and cell proliferation of the AT-MSCs were studied on the days 5, 7, and 11 by an inverted microscope and MTT assay, respectively. To evaluate the stemness state of the cells, Oct4 expression was measured using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Also, differentiation potential of AT-MSCs toward adipogenic and osteogenic lineages was assessed. All tests were done in triplicate.

Differentiation potential of adipose tissue‐derived mesenchymal stem cells into germ cells with and without growth factors

“…The pattern of CD marker expression was similar to the MSCs. CD90 and CD44 were overexpressed in the isolated cells, but the expression of the CD31 and CD45 was low, as found in the previous study (Eliyasi Dashtaki et al, 2018).…”

“…To increase the efficiency of the differentiation, studies are performed to provide a suitable niche for stem cells. In our previous study, three different doses of EGF, GDNF and LIF were added to the culture of AT-MSCs and it was observed that the viability of the cells in the presence of EGF (10 ng/ml), GDNF (5 ng/ml) and LIF (5 ng/ml) was increased (Eliyasi Dashtaki et al, 2018). As we previously reported that the mice transplanted with AT-MSCs supplemented with the aforementioned growth factors showed more expression of Mvh and Gcnf compared to the mice transplanted with cells, cultured in medium without growth factors (Eliyasi Dashtaki et al, 2020).…”

“…The 3rd passage of the cells was utilised for tests. The expression of specific MSCs CD markers (CD90 and CD44) and the non-expression of hematopoietic stem cell and endothelial cell CD markers (CD31 and CD45) were evaluated by flow cytometry, as explained in our previous study (Eliyasi Dashtaki et al, 2018).…”

Effects of clinical medications on male fertility and prospects for stem cell therapy

“…Frontiers in Cell and Developmental Biology frontiersin.org (Abdelaziz et al, 2019;Elbaghdady et al, 2018;Sherif et al, 2018), and testicular injection also has an improvement effect (Hsiao et al, 2015;SM et al, 2017). ADSCs and AFSCs can reduce oxidative stress in the testes and promote sperm production, (Eliyasi et al, 2020;Ibrahim et al, 2021;Qian et al, 2020;Siregar et al, 2021), and can improve the damage of chemotherapy drugs to testes (Meligy et al, 2019). Intratesticular injection of ADSCs can improve the spermatogenesis of azoospermia (Ganjibakhsh et al, 2022;Hajihoseini et al, 2018;Karimaghai et al, 2018), while xenotransplantation of human umbilical cord blood derived MSCs into azoospermia mice also has the same effect (Abd et al, 2017).…”

The determination of epidermal growth factor in Edible bird's nest by enzyme-linked immunosorbent assay

“…The molecular weight of EGF is about 6000 Da, and the isoelectric point of EGF is 4.6 [13]. EGF participated in various physiological activities, such as cell division, proliferation, and migration [14][15][16][17]. It could promote the growth of epidermal cells, epithelial tissue regeneration, and wound healing, which had been widely used in the clinical treatment of skin wounds [18][19][20].…”