Optimization of human mesenchymal stem cell manufacturing: the effects of animal/xeno-free media

Abstract

Due to their immunosuppressive properties, mesenchymal stem cells (MSC) have been evaluated for the treatment of immunological diseases. However, the animal-derived growth supplements utilized for MSC manufacturing may lead to clinical complications. Characterization of alternative media formulations is imperative for MSC therapeutic application. Human BMMSC and AdMSC were expanded in media supplemented with either human platelet lysates (HPL), serum-free media/xeno-free FDA-approved culture medium (SFM/XF), or fetal bovine serum (FBS) and the effects on their properties were investigated. The immunophenotype of resting and IFN-γ primed BMMSC and AdMSC remained unaltered in all media. Both HPL and SFM/XF increased the proliferation of BMMSC and AdMSC. Expansion of BMMSC and AdMSC in HPL increased their differentiation, compared to SFM/XF and FBS. Resting BMMSC and AdMSC, expanded in FBS or SFM/XF, demonstrated potent immunosuppressive properties in both non-primed and IFN-γ primed conditions, whereas HPL-expanded MSC exhibited diminished immunosuppressive properties. Finally, IFN-γ primed BMMSC and AdMSC expanded in SFM/XF and HPL expressed attenuated levels of IDO-1 compared to FBS. Herein, we provide strong evidence supporting the use of the FDA-approved SFM/XF medium, in contrast to the HPL medium, for the expansion of MSC towards therapeutic applications.

Pathogen reduction through additive-free short-wave uv light irradiation retains the optimal efficacy of human platelet lysate for the expansion of human bone marrow mesenchymal stem cells

“…This result was in agreement with other studies that did not observe any significant difference between those two supplements in terms of immunosuppressive properties of BM-hMSCs [12,13,47]. However, some studies showed that BM-hMSCs cultured in hPL displayed lesser properties of T-cell proliferation inhibition than BM-hMSCs cultured in FBS [11,48,49]. In our study, an increase of T-cell proliferation inhibition was even obtained with PRhPL but only at the 1:5 ratio.…”

Directed differentiation of human iPSC into insulin producing cells is improved by induced expression of PDX1 and NKX6.1 factors in IPC progenitors

“…Additional concerns involve the contamination with adventitious agents that may result in animal-to-human pathogen transmission in the course of therapy. As a consequence, there is a high demand from regulatory bodies such as Food and Drug Administration (FDA) and the European Medicines Agency (EMA) to put restrictions on the application of animal products in cell-based therapeutics [22, 23]. For this reason, our aim was to establish efficient and reliable protocols based on defined components.…”

Heterogeneous Niche Activity of Ex-Vivo Expanded MSCs as Factor for Variable Outcomes in Hematopoietic Recovery

“…These findings suggest that the CFU-F content of ex-vivo cultured MSCs, unlike the inherent clonal differences among MSC subsets in BM, can be readily switched by extrinsic signals, and that it can be a barometer for niche activities for a variety of culture conditions including the xeno-free culture conditions currently being developed [53, 54]. …”