Abstract
IntroductionFetal bovine serum (FBS) is an animal product used as a medium supplement. The animal origin of FBS is a concern if cultured stem cells are to be utilized for human cell therapy. Therefore, a substitute for FBS is desirable. In this study, an industrial, xeno-free, pharmaceutical-grade supplement for cell culture (SCC) under development at Grifols was tested for growth of human mesenchymal stem cells (hMSCs), cell characterization, and differentiation capacity.MethodsSCC is a freeze-dried product obtained through cold-ethanol fractionation of industrial human plasma pools from healthy donors. Bone marrow-derived hMSC cell lines were obtained from two commercial suppliers. Cell growth was evaluated by culturing hMSCs with commercial media or media supplemented with SCC or FBS. Cell viability and cell yield were assessed with an automated cell counter. Cell surface markers were studied by indirect immunofluorescence assay. Cells were cultured then differentiated into adipocytes, chondrocytes, osteoblasts, and neurons, as assessed by specific staining and microscopy observation.ResultsSCC supported the growth of commercial hMSCs. Starting from the same number of seeded cells in two consecutive passages of culture with medium supplemented with SCC, hMSC yield and cell population doubling time were equivalent to the values obtained with the commercial medium and was consistent among lots. The viability of hMSCs was higher than 90%, while maintaining the characteristic phenotype of undifferentiated hMSCs (positive for CD29, CD44, CD90, CD105, CD146, CD166 and Stro-1; negative for CD14 and CD19). Cultured hMSCs maintained the potential for differentiation into adipocytes, chondrocytes, osteoblasts, and neurons.ConclusionsThe tested human plasma-derived SCC sustains the adequate growth of hMSCs, while preserving their differentiation capacity. SCC can be a potential candidate for cell culture supplement in advanced cell therapies.
Highlights
Fetal bovine serum (FBS) is an animal product used as a medium supplement
Starting from the same number of seeded cells in two consecutive passages of culture with medium supplemented with supplement for cell culture (SCC), human mesenchymal stem cell (hMSC) yield and cell population doubling time were equivalent to the values obtained with the commercial medium and was consistent among lots
The viability of hMSCs was higher than 90%, while maintaining the characteristic phenotype of undifferentiated hMSCs
Summary
The animal origin of FBS is a concern if cultured stem cells are to be utilized for human cell therapy. HMSCs have a low immunogenicity – which can help to improve allogenic transplantation and avoid immune rejection, one of the main complications of stem cell use in cell therapy [3,4,5,6,7,8]. These characteristics make hMSCs of great interest for use in regenerative medicine therapies, as well. The need for a stable, standard, and safe in vitro culture medium is one of the major shortcomings of hMSC use in advanced therapies.
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