Enhanced Cell Growth of Adipocyte-Derived Mesenchymal Stem Cells Using Chemically-Defined Serum-Free Media

Abstract

The multipotency and anti-inflammatory effects of mesenchymal stem cells (MSCs) make them attractive for cell therapy in regenerative medicine. A large number of MSCs is required for efficient therapy owing to the low homing efficiency of MSCs to target sites. Furthermore, owing to limitations in obtaining sufficient amounts of MSCs, in vitro expansion of MSCs that preserves their differentiation and proliferative potential is essential. The animal factor included in culture media also limits clinical application. In this study, adipose-derived MSCs showed a significantly higher proliferation rate in STK2, a chemically-defined medium, than in DMEM/FBS. The expression of MSC surface markers was increased in the culture using STK2 compared to that using DMEM/FBS. Tri-lineage differentiation analyses showed that MSCs cultured in STK2 were superior to those cultured in DMEM/FBS. In addition, MSCs cultured in STK2 showed a reduced senescence rate, small and homogenous cell size, and were more genetically stable compared to those cultured in DMEM/FBS. Furthermore, secretome analysis showed that the expression of factors related to proliferation/migration, anti-inflammation, and differentiation were increased in STK2 culture medium compared to DMEM/FBS. Taken together, these results suggest that culture using STK2 medium offers many advantages through which it is possible to obtain safer, superior, and larger numbers of MSCs.

The Secretome Derived From Mesenchymal Stromal Cells Cultured in a Xeno-Free Medium Promotes Human Cartilage Recovery in vitro

“…hBMSCs isolated and cultured in XFS showed a higher and more stable proliferation rate than FBS-hBMSCs, suggesting that the XFS culture can enable the efficient production of therapeutic cells in a shorter time. These data are consistent with those reported for adipose-derived MSCs and human dental pulp stem cells cultured in chemically defined xenofree media (Lee et al, 2017;Mochizuki and Nakahara, 2018). The maintenance of the typical MSC-phenotype according to the ISCT criteria (Dominici et al, 2006) underpins the use of this XFS culture system for MSC isolation.…”

Limited Potential or Unfavorable Manipulations? Strategies Toward Efficient Mesenchymal Stem/Stromal Cell Applications

“…As an alternative, human serum or human platelet lysate can be used for MSCs' expansion (Mannello and Tonti, 2007;Bieback, 2013;Hemeda et al, 2014). Chemically defined xeno-free media could provide acceptable cell growth and elimination of the risk of carrying over pathogens (Spees et al, 2004), but the evaluation of functional characteristics of the cells cultivated in such media has to be improved (Lee et al, 2017).…”

Mesenchymal Stromal Cells: From Discovery to Manufacturing and Commercialization

“…Ideally, a suitable FBS substitute should present defined composition, reduced risk of contamination, low costs, easy availability, and extended shelf life [ 73 ]. Some studies have reported the use of a stirred tank bioreactor for MSC expansion under xenogeneic- (xeno-) free conditions with the use of human serum [ 39 ] and chemically defined culture medium [ 74 , 75 ]. The first FDA approved commercial xeno-free culture medium was StemPro MSC SFM (Invitrogen).…”