Human Alternatives to Fetal Bovine Serum for the Expansion of Mesenchymal Stromal Cells from Bone Marrow

Abstract

Mesenchymal stromal cells (MSCs) are promising candidates for novel cell therapeutic applications. For clinical scale manufacturing, human factors from serum or platelets have been suggested as alternatives to fetal bovine serum (FBS). We have previously shown that pooled human serum (HS) and thrombin-activated platelet releasate in plasma (tPRP) support the expansion of adipose tissuederived MSCs. Contradictory results with bone marrow (BM)-derived MSCs have initiated a comprehensive comparison of HS, tPRP, and pooled human platelet lysate (pHPL) and FBS in terms of their impact on MSC isolation, expansion, differentiation, and immunomodulatory activity. In addition to conventional Ficoll density gradient centrifugation, depletion of lineage marker expressing cells (RosetteSep) and CD271 1 sorting were used for BM-MSC enrichment. Cells were cultured in medium containing either 10% FBS, HS, tPRP, or pHPL. Colony-forming units and cumulative population doublings were determined, and MSCs were maximally expanded. Although both HS and tPRP comparable to FBS supported isolation and expansion, pHPL significantly accelerated BM-MSC proliferation to yield clinically relevant numbers within the first two passages. MSC quality and functionality including cell surface marker expression, adipogenic and osteogenic differentiation, and immunosuppressive action were similar in MSCs from all culture conditions. Importantly, spontaneous cell transformation was not observed in any of the culture conditions. Telomerase activity was not detected in any of the cultures at any passage. In contrast to previous data from adipose tissue-derived MSCs, pHPL was found to be the most suitable FBS substitute in clinical scale

Culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in different media

“…al 2012. However, to avoid the potential risk of exposing human patients to cells cultivated in the presence of FBS, some investigators have searched for a substitute supplement (Bieback et al 2009) because, according to the results of Pal et al (2009), the presence of serum is essential for maintaining and expanding mesenchymal stem cell cultures. In the present work, despite samples coming from adipose tissue, they showed greater cellularity in DMEM supplemented with FBS in the first two weeks, but afterwards, there is no longer any difference in relation to grown in AIM-V medium without FBS, suggesting that this can be an alternative for the culture of cells destined for transplantation.…”

Inactivated human platelet lysate with psoralen: a new perspective for mesenchymal stromal cell production in Good Manufacturing Practice conditions

“…Serum-free medium is unable to promote MSC expansion unless several recombinant human growth factors (GF) such as platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), transforming growth factor (TGF)-β, and epidermal growth factor (EGF) are added to the medium. However, 1) defining the optimal amount of GF is difficult, 2) only a few GFs are licensed for therapeutic use, 3) recombinant Human-GFs are expensive, and 4) isolated GF cannot replace the different physiological functions of FBS (15)(16)(17).…”

Effect of fetal bovine serum on mineralization in silk fibroin scaffolds

“…The use of human serum, however, makes sense only if enough autologous serum is available. Due to the limited availability and high variability within different donors, human serum is not a reasonable option for large-scale cell cultures in clinical applications [46] and therefore it is highly recommended to look for serum-free alternatives for in vitro cultures.…”