Human adipose derived stroma/stem cells grow in serum-free medium as floating spheres.
With the goal of obtaining clinically safe human adipose-derived stroma/stem cells (ASC) and eliminating the use of serum, we have developed a new culture system that allows the expansion of ASC as spheres in a defined medium. These spheres can be passaged several times. They are not only aggregated cells but rather originate from single cells as clonal spheres can be obtained after seeding at very low density and reform clonal spheres after dissociation. These spheres can also revert to monolayer growth when plated in medium containing human plasma and even generate fibroblast-like colonies (CFU-f). Under several differentiation-specific media, spheres-derived ASC maintain their capacity to differentiate into osteoblasts, endothelial cells and adipocytes. These results indicate that human ASC can be maintained in a serum-free 3D culture system, which is of great interest for the expansion in bioreactors of autologous ASC and their use in clinical trials.
Université de Toulouse, UPS, UMR5241 Métabolisme, Plasticité et Mitochondrie, BP 84 225, F-31 432 Toulouse, France.
C
C
Dromard
P
P
Bourin
M
M
André
S
S
De Barros
L
L
Casteilla
V
V
Planat-Benard
D017667:Adipocytes / Q000166:cytology*; D000273:Adipose Tissue / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D019070:Cell Lineage; D016895:Culture Media, Serum-Free; D005434:Flow Cytometry; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D016895:Culture Media, Serum-Free