Human marrow mesenchymal stem cell culture: serum-free medium allows better expansion than classical alpha-MEM medium.
The expansion of mesenchymal stem cells (MSCs) strongly depends on the culture conditions and requires medium supplemented with 10-20% fetal calf serum (FCS) to generate relevant numbers of cells. However, the presence of FCS is a major obstacle for their clinical use. Therefore, we have evaluated the capacity of expansion of MSC in a commercial serum-free medium (UC) supplemented with a serum substitute (ULTROSER) in comparison with a classical medium alpha-MEM containing 15% FBS. Bone marrow-mononuclear cells collected from 12 volunteer healthy donors were expanded in two different culture media. MSCs isolated in the both media were morphologically similar and expressed identical phenotypic markers. After the primoculture (P0) and one passage, we obtained significantly more MSC and CFU-F progenitors in UC medium than in alphaMEM. Their multipotentiality was preserved during culture, as well as their capacity to support haematopoiesis. In conclusion, our observations strongly suggest that UC is an optimal medium for ex vivo expansion of MSC: it allows a better cell expansion, preserves cell multipotentiality, reduces the culture period and contains low concentration of serum substitute. This medium seems suitable for clinical scale expansion of MSC.
Meuleman
Nathalie
N
Department of Experimental Hematology, Institut Jules Bordet, Centre des Tumeurs de l'Université Libre de Bruxelles, Brussels, Belgium. nathalie.meuleman@bordet.be
Tondreau
Tatiana
T
Delforge
Alain
A
Dejeneffe
Marielle
M
Massy
Martine
M
Libertalis
Mark
M
Bron
Dominique
D
Lagneaux
Laurence
L
D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D006410:Hematopoiesis / Q000502:physiology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D009930:Organic Chemicals
D016895:Culture Media, Serum-Free; D009930:Organic Chemicals; C420642:alpha minimal essential medium