Which Culture System Is better for Chondrogenesis of Adipose-Derived Stem Cells ; Pellet or Micromass?

Background and Aims: The current study was conducted to compare the expression levels of collagen type Π and X during chondrogenesis of human adipose-derived mesenchymal stem cells (hADMSCs) pellet and micromass cultures.  Materials and Methods: Extracted hADMSCs were cultured until three passages and then transferred to pellet and micromass cultures in the experimental groups of day ۷ and day۱۴. For pellet and micromass cultures, aliquots of ۵×۱۰۵ cells/ml were centrifuged and respectively cultured in the conical tubes and droplets (۱۲.۵ µl) of the ۲۴-well plates containing chondrogenic medium. Realtime-polymerase chain reaction technique was performed for gene expression levels. Results: Increased expression of collagen type Π was shown in micromass day۱۴ compared to micromass day ۷, pellet day ۱۴ (p<۰.۰۱) and pellet day ۷ (p<۰.۰۰۱). Also, an increased expression of collagen type Π was seen in micromass day ۷ and pellet day ۱۴ compared to pellet day ۷ (p< ۰.۰۵). Expression of collagen type X increased in pellet day ۱۴ compared to micromass on days ۷ and ۱۴ (p<۰.۰۰۱, p<۰.۰۱) and pellet day۱۴ compared to pellet day۷ (p< ۰.۰۵). An increased expression of collagen type X was shown in pellet day ۷ compared to micromass on days ۷ and ۱۴ (p<۰.۰۵). Conclusions: According to the results, higher expression of collagen type Π and lower expression of collagen type X in micromass cultures that are prepared by cell suspension play a better role during cellular condensation that leads to the formation of large nodules exhibiting cartilage-like morphology, suggests a higher efficiency for micromass cultures.

Keywords:

Chondrogenesis , Collagen , Stem cell