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A new clinical‐scale serum‐free xeno‐free medium efficient in ex vivo amplification of mesenchymal stromal cells does not support mesenchymal stem cells
Author(s) -
Gerby Sandie,
Attebi Esther,
Vlaski Marija,
Ivanovic Zoran
Publication year - 2017
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.13902
Subject(s) - mesenchymal stem cell , immunophenotyping , ex vivo , biology , stromal cell , stem cell , cell culture , population , flow cytometry , immunology , microbiology and biotechnology , in vivo , cancer research , medicine , genetics , environmental health
BACKGROUND We evaluated a new serum‐free, xeno‐free medium (Xuri, GE HealthCare) in ex vivo cultures for amplification of mesenchymal stromal cells (MStroC) in comparison with classical culture supplemented with fetal calf serum and basic fibroblast growth factor. STUDY DESIGN AND METHODS MStroC and mesenchymal stem cell (MSC) proliferative capacities were studied in bulk cultures and single‐cell cultures with assay of secondary replating capacity of individual clones. Flow‐cytometric phenotype analysis and proliferative history analysis were also performed. RESULTS In cultures initiated with previously amplified and cryopreserved MStroC from human marrow, Xuri medium enabled a total cell expansion fold comparable to one obtained in control fetal calf serum (FCS)‐supplemented culture. However, both the number and the proliferative capacity of colony‐forming unit–fibroblast were greatly reduced in Xuri medium cultures. This is even more evident in single‐cell cultures, where, in rare positive wells, only several cells were found in Xuri cultures, compared to abundant cell content in FCS and α‐minimal essential medium cultures. Replating these single‐cell clones in secondary cultures (FCS in both cases) revealed a total exhaustion of MSC proliferative capacity after Xuri primary culture. CONCLUSION Since in both conditions after a 7‐day bulk culture, similar immunophenotype and proliferative history were found when the standard MSC immunophenotype panel was employed, the loss of proliferative capacity in Xuri medium shows that it cannot maintain functional MSC population. This is a drastic example showing that the real MSC activity can be completely unrelated to the immunophenotype considered as MSC phenotype.