Xenogen-free media provide variable equine mesenchymal stromal cell expansion after a 7-day culture period
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Updated: Jan 16, 2026
Production and Administration of Therapeutic Mesenchymal Stem/Stromal Cell MSC Spheroids Primed in 3-D Cultures Under Xeno-free Conditions
Published on: March 18, 2017
Objective:
To determine the xenogen-free serum source that provides the greatest number of live equine mesenchymal stromal cells (MSCs) while maintaining the MSC phenotype.
Methods:
Equine bone marrow-derived MSCs from 8 horses were cultured for 7 days in media containing one of the following serum treatments: 10% xenogeneic serum, 10% or 20% commercial allogeneic equine serum, 10% autologous serum, 10% equine pooled platelet lysate (PPL), or a staged media reduction of xenogeneic media. Live cell numbers, MSC viability, and MSC immunophenotype were compared.
Results:
The use of 10% commercial allogeneic equine serum in media resulted in a significant decrease in live MSC count compared to xenogeneic serum (P = .01; 95% CI, -244,766 to -48,038). Autologous serum, staged reduction, and PPL had no significant difference in live MSC number collected at day 7 as compared to xenogeneic serum. There were no significant differences in cell count due to the horse's age group. Viability was significantly greater using PPL than all other xenogen-free serums across the < 10-year and ≥ 10-year age groups. Mesenchymal stromal cells from all treatments were high in CD44, CD90, and major histocompatibility complex (MHC) I expression and low in CD45 and MHC II expression. Significant differences in CD44, CD45, CD90, MHC I, and MHC II expression levels were seen across treatment and age analysis.
Conclusions:
The use of PPL for MSC media in the final 7 days of culture allows for adequate expansion and viability of MSCs and maintenance of MSC immunophenotypic markers.
Clinical Relevance:
MSCs can be cultured in nonxenogeneic media for 7 days prior to harvest.
