Different doses of bone morphogenetic protein 4 promote the expression of early germ cell-specific gene in bone marrow mesenchymal stem cells

Abstract

Bone morphogenetic protein (BMP)-4 has a crucial role on primordial germ cells (PGCs) development in vivo which can promote stem cell differentiation to PG-like cells. In this study, we investigated the expression of Mvh as one of the specific genes in primordial germ cells after treatment with different doses of BMP4 on bone mesenchymal stem cells (BMSCs)-derived PGCs. Following isolation of BMSCs from male mouse femur and tibia, cells were cultured in medium for 72 h. Passage 4 murine BMSCs were characterized by CD90, CD105, CD34, and CD45 markers and osteo-adipogenic differentiation. Different doses of BMP4 (0, 0.01, 0.1, 1, 5, 25, 50, and 100 ng/ml) were added to BMSCs for PGCs differentiation during 4-days culture. Viability percent, proliferation rates, and expression of Mvh gene were analyzed by RT-qPCR. Data analysis was done with ANOVA test. CD90(+), CD105(+), CD34(-), and CD45(-) BMSCs were able to differentiate to osteo-adipogenic lineages. The results revealed that proliferation rate and viability percent were raised significantly (p ≤ 0.05) by adding 1, 5, 25 ng/ml of BMP4 and there were decreased to the lowest rate after adding 100 ng/ml BMP4 (p ≤ 0.05). There were significant up regulation (p ≤ 0.05) in Mvh expression between 25, 50, and 100 ng/ml BMP4 with other doses. So the selective dose of BMP-4 for treatment during 4-day culture was 25 ng/ml. The results suggest that addition of 25 ng/ml BMP4 had the best effects based on gene-specific marker expression.

Influence of Activin A Supplementation During Human Embryonic Stem Cell Derivation on Germ Cell Differentiation Potential

“…Kee and colleagues observed that adding BMP4 at 1 ng/mL, 10 ng/mL and 100 ng/mL corresponded with a significant rise in expression of VASA [2]. Directed differentiation of bone marrow mesenchymal stem cells towards germ cells was also accompanied by significant upregulation of VASA when the BMP4 concentration was increased from 25 ng/mL to 100 ng/mL [37]. Similarly, we also observed that BMP4 had a dose-dependent effect on PGC differentiation as 50 ng/mL and 100 ng/mL BMP4 significantly upregulated …”

“…PGC-directed differentiation of bone marrow mesenchymal stem cells also showed a BMP4-associated dose dependent increase in VASA expression where 100 ng/mL proved to be the best concentration compared to 25 ng/mL and 50 ng/mL [37]. A cocktail of BMPs, including BMP4, BMP7, and BMP8b supplemented at 100 ng/mL has also proven to be highly beneficial in germ cell differentiation of hESCs compared to 1 ng/mL and 10 ng/ mL of BMP4 [2].…”

Pericyte‐derived bone morphogenetic protein 4 underlies white matter damage after chronic hypoperfusion

“…This finding was inconsistent with other studies that demonstrated no effect 16, 57 or even a positive effect 43 of BMP4 on OPC proliferation. However, recent studies have suggested that the effects of BMP4 on stem cell proliferation are dose‐dependent; low dose BMP4 increases proliferation, but high dose BMP4 decreases proliferation of mesenchymal stem cells 53 or primordial germ cells 31. Considering that OPC proliferation is also affected by many factors, BMP4 may have dose‐dependent and opposite effects on OPC proliferation.…”

Inductive Role of Sustentacular Cells (Sertoli Cells) Conditioned Medium on Bone Marrow Derived Mesenchymal Stem Cells

“…The cells were incubated in 5 mg/ml fluorescein isothiocyanate (FITC) conjugated antibodies. FITC mouse IgG2A (11-4724, eBioscience, UK) and Rat IgG1 (ab18412, abcam, USA) were used as isotype controls (Mazaheri et al, 2011;Rastegar et al, 2013). Sustentacular cells isolation.…”