Abstract
BackgroundMesenchymal stem cells (MSCs) show promising characteristics for their use in advanced therapy medicinal products. However, there are some unresolved concerns, such as the use of animal components for their expansion.In this study we assessed the suitability of a xeno-free supplement for cell culture (SCC) derived from human plasma, to culture and expand human MSCs (hMSCs) from different origins. Characteristics of viable cultured hMSCs such as genetic stability, phenotype and multipotentiality were qualitatively evaluated.MethodshMSCs from adipose tissue (AT), bone marrow (BM) and umbilical cord (UC) and supplier sources (commercial/non-commercial) were used. After hMSCs expansion in a xeno-free medium, classical hMSCs markers were studied by immunocytochemistry, and genetic stability was tested by classic karyotyping. The capacity of hMSCs to differentiate into adipogenic, osteogenic, and chondrogenic cells in differentiation media was assessed using different staining. Different lots of SCC were used to assure consistency between batches.ResultsAll hMSCs tested maintained their morphology and adherence to plastic during their expansion, and preserved their genetic stability, phenotype and differentiation potential. No differences were observed when using different lots of SCC. Moreover, the proliferation rate, evaluated as population doubling time (PDT) of commercial BM and AT hMSCs, was higher in the xeno-free medium than in the control media provided by the suppliers of the cells (PDT of 4.6 for BM-hMSC and 6.4 for AT-hMSC in xeno-free medium, and 7.0 and 14.7 respectively in the commercial media). UC-hMSCs PDT was similar in all the media tested. When using non-commercial BM-hMSCs, PDT was lower in the xeno-free medium, but reverted to the control level with the addition of growth factors.ConclusionsSCC-containing medium can be a feasible xeno-free alternative to expand hMSCs for advanced therapies.
Highlights
Mesenchymal stem cells (MSCs) show promising characteristics for their use in advanced therapy medicinal products
Results hMSc growth in xeno-free conditions bone marrow (BM)-Human mesenchymal stem cell (hMSC), AThMSCs and umbilical cord (UC)-hMSCs could be expanded in xeno-free medium, with (PL-Medium) and without (XF-Medium) platelet lysate
The addition of platelet lysate into the xeno-free medium did not produce significant changes in the population doubling time (PDT) of the commercial hMSCs lines (BM-hMSC = 4.8, adipose tissue (AT)-hMSC = 6.2, UC-hMSC = 1.9)
Summary
Mesenchymal stem cells (MSCs) show promising characteristics for their use in advanced therapy medicinal products. One of the main concerns is related to the use of fetal bovine serum (FBS) as a supplement in cell culture medium to expand hMSCs. The introduction of animal derivatives into human cell cultures is not recommended since animal proteins can become associated with MSCs, and induce an immune rejection to the host [4]. The introduction of animal derivatives into human cell cultures is not recommended since animal proteins can become associated with MSCs, and induce an immune rejection to the host [4] Another drawback is the lack of lot-to-lot consistency of the FBS and its limited source [5,6,7]. The presence of animal components is discouraged in the culture media used to expand hMSC for therapy [4]
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