Abstract
Culture medium of mesenchymal stromal cells (MSCs) is usually supplemented with either human platelet lysate (HPL) or fetal calf serum (FCS). Many studies have demonstrated that proliferation and cellular morphology are affected by these supplements – it is therefore important to determine if they favor outgrowth of different subpopulations and thereby impact on the heterogeneous composition of MSCs. We have isolated and expanded human bone marrow-derived MSCs in parallel with HPL or FCS and demonstrated that HPL significantly increases proliferation and leads to dramatic differences in cellular morphology. Remarkably, global DNA-methylation profiles did not reveal any significant differences. Even at the transcriptomic level, there were only moderate changes in pairwise comparison. Furthermore, the effects on proliferation, cytoskeletal organization, and focal adhesions were reversible by interchanging to opposite culture conditions. These results indicate that cultivation of MSCs with HPL or FCS has no systematic bias for specific cell types.
Highlights
Fetal calf serum is usually considered as the gold standard for MSCs culture
When cells that were initially cultured in FCS were transferred to HPL for two days, they adopted similar focal adhesions as those that were continuously cultured in HPL. These results demonstrate that the differences in proliferation rate, cellular morphology, and cell-biomaterial interaction are reversible by interchanging culture conditions
Precise definition of cell preparations is essential for reproducible results – and this applies to MSCs intended for therapeutic application
Summary
Fetal calf serum is usually considered as the gold standard for MSCs culture. there is high variation between FCS batches and it involves the risk of transmitting bovine infections or initiation of xenogeneic immune responses. Apart from the regulatory concerns and impact on proliferation it may be even more important to understand the biological sequel of these supplements on MSC preparations[8]. It has been demonstrated that culture conditions with either HPL or FCS give rise to MSCs with different morphological features[9, 10]. Given the heterogeneous composition of MSCs it may be anticipated that specific subpopulations are selected – or at least favored - by one or the other culture supplement. The highly reproducible and quantitative differences in DNA-methylation (DNAm) patterns – which are apparently directly involved in cell type specification – provide an ideal basis for molecular characterization and definition of cell types[12]. We have isolated and continuously cultured MSCs in parallel with HPL and FCS to directly compare their DNAm patterns and molecular characteristics
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
