Biological Features of Human Bone Marrow Stromal Cells (hBMSC) Cultured with Animal Protein-Free Medium—Safety and Efficacy of Clinical Use for Neurotransplantation

Abstract

The donor cell culture in animal serum-free medium is quite important for the clinical application of cell transplantation therapy. This study was aimed to test the hypothesis that the human bone marrow stromal cells (hBMSC) expanded with fetal calf serum (FCS)-free, platelet lysate (PL)-containing medium retain their biological features favoring central nervous system regeneration. The hBMSC were cultured with 5% PL or 10% FCS. Their phenotypes were analyzed with flow cytometry, and their production of growth factors was quantified with enzyme-linked immunosorbent assay. Their capacity of neural differentiation was verified by immunocytochemistry. There was no significant difference in morphology and cell surface marker between the hBMSC-FCS and hBMSC-PL. Both of them were positive for CD44, CD90, CD105, and CD166 and were negative for CD34, CD45, and CD271. The production of human brain-derived neurotrophic factor, human hepatocyte growth factor, human β-nerve growth factor, and human platelet-derived growth factor-BB did not differ between the two groups, although the hBMSC-PL produced significantly more amount of TGF-β1 than the hBMSC-FCS. There was no significant difference in their in vitro differentiation into the neurons and astrocytes between the two groups. The hBMSC expanded with PL-containing medium retain their biological capacity of neural differentiation and neuroprotection. The PL may be a clinically valuable and safe substitute for FCS in expanding the hBMSC for cell therapy.

Feasibility and Efficiency of Human Bone Marrow Stromal Cell Culture with Allogeneic Platelet Lysate‐Supplementation for Cell Therapy against Stroke

“…In our previous study, no significant differences were observed between hBMSCs cultured using 5% PL and using 10% FCS [17]. In the present study, although the concentration of PL was changed to 10%, we confirmed that the hBMSCs were identical for surface markers.…”

“…In contrast, our previous report showed that the concentration of human BDNF, which was derived from PL, markedly decreased in the medium after hBMSC culture. These results strongly suggested that the cultured hBMSCs may consume human BDNF for their survival and proliferation [17]. In the present study, we found that the content of these growth factors in each PL was relevant to the amount of platelets in original PC.…”

Advancing biomaterials of human origin for tissue engineering

“…It is suggested that cell culture with PL supplementation does not change the expression of surface markers on cultured cells compared to that in cultures supplemented with FBS [634]. Most importantly, these cells retain their biological features, such as their growth and differentiation potential favoring target tissue regeneration [634,635].…”

“…It is suggested that cell culture with PL supplementation does not change the expression of surface markers on cultured cells compared to that in cultures supplemented with FBS [634]. Most importantly, these cells retain their biological features, such as their growth and differentiation potential favoring target tissue regeneration [634,635]. In recent years, cumulative evidence has strongly indicated that human PL-supplemented media not only preserve cell phenotypes and shorten culture time by increasing the cells' growth rate but also maintain the cells' multilineage differentiation capacity and can give rise to osteoblasts, chondrocytes, adipocytes, neurons and astrocytes, among others [630,634–638].…”

“…Most importantly, these cells retain their biological features, such as their growth and differentiation potential favoring target tissue regeneration [634,635]. In recent years, cumulative evidence has strongly indicated that human PL-supplemented media not only preserve cell phenotypes and shorten culture time by increasing the cells' growth rate but also maintain the cells' multilineage differentiation capacity and can give rise to osteoblasts, chondrocytes, adipocytes, neurons and astrocytes, among others [630,634–638]. Interestingly, accompanied by a more compact colony formation and a more spindle-shaped morphology, PL-cultured BMSCs [639] and ADSCs [640] remained significantly smaller in size than did FBS-cultured cells.…”