Validation of reference and identity-defining genes in human mesenchymal stem cells cultured under unrelated fetal bovine serum batches for basic science and clinical application

The molecular profile of human mesenchymal stem cells (MSCs) have emerged as a key factor in defining their identity. Nevertheless, the effect of fetal bovine serum (FBS) batches or origin on MSC molecular signature has been neglected. In this frame, chemical fingerprint of FBS batches from unrelate...

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Bibliographic Details
Published in:Stem cell reviews and reports 2018-12, Vol.14 (6), p.837-846
Main Authors: Banfi, Federica, Colombini, Alessandra, Perucca Orfei, Carlotta, Parazzi, Valentina, Ragni, Enrico
Format: Article
Language:English
Subjects:
Animals
Antigens, Differentiation - biosynthesis
Bioinformatics
Bone marrow
Bone Marrow Cells - cytology
Bone Marrow Cells - metabolism
Cattle
Cell Culture Techniques - standards
Culture Media - standards
Data processing
Fetal Blood - cytology
Fetal Blood - metabolism
Fetal calf serum
Gene expression
Humans
Male
Mesenchymal stem cells
Mesenchymal Stem Cells - cytology
Mesenchymal Stem Cells - metabolism
Mesenchyme
Middle Aged
Reference Standards
Serum
Stem cells
Transcription
Umbilical cord
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Summary:The molecular profile of human mesenchymal stem cells (MSCs) have emerged as a key factor in defining their identity. Nevertheless, the effect of fetal bovine serum (FBS) batches or origin on MSC molecular signature has been neglected. In this frame, chemical fingerprint of FBS batches from unrelated countries showed strong correlation between chemical composition and country of origin. Thus, the aim of this study was to evaluate in stem cells isolated from bone marrow (BMMSCs) and umbilical cord-blood (CBMSCs) the effects of independently collected FBS batches on both twelve commonly used reference genes (RGs) and a selected panel of thirty-eight genes crucial for MSC definition in both research and clinical settings. Gene expression stability was estimated comparing the outcomes of two applets: geNorm and NormFinder. The bioinformatics analysis emphasized that, in a panorama of general balance, few RG candidates (YWHAZ/UBC for BMMSCs, RPLP0/EF1A for CBMSCs and EF1A/TBP for both MSCs scored together) showed superior stability. In addition, a wider study on genes involved in differentiation/proliferation/stemness processes, often used to define MSC potency, showed that these genes exhibited no major transcriptional modulation after treatment with different FBS, and allowed the identification of genes strongly discriminating between BM- and CBMSC populations. Therefore, in conclusion, FBS origin does not dramatically impact the general molecular profile of MSCs, although we could identify validated candidates able to allow more reliable comparison of data regarding MSC identity and potency and obtained by research laboratories and clinical manufacturers using different sera.
ISSN:2629-3269
2629-3277
DOI:10.1007/s12015-018-9822-0