Supplementation of TGF-Beta3 in Low Serum Media Promotes Chondrogenesis of BMSCs

Tissue  engineering has  emerged   as  a  new  promising  field  that  allow  in  vitro construction of whole  transplantable tissue.  Recently,  bone  marrow  stem  cells (BMSCs) fulfil the  requirements as appropriate cell  source  that  is renewable for cartilage  tissue  engineering since  they  were  able  to form  hyaline-like  cartilage in vitro and  in vivo  when  cultured  in media  supplemented with specific growth factors. This study aimed  to examine  the potential  of BMSCs chondrogenesis in vitro and in vivo.  BMSCs were isolated  from sheep  and divided  into a test group and  control  group.  Control  group  was  cultured  in medium  supplemented with 10% fetal bovine  serum (FBS). Test group was cultured  in medium  supplemented with 1% FBS, 1% ITS, 5ng/mL TGF-ß3, 50ng/mL IGF-1, 40 ng/mL L-proline, 100 nM dexamethasone and  50 μg/mL ascorbic  acid-2  phosphate. The experiment was carried out on athymic nude mice. Cell aggregates were formed in test group and indicated the early chondrogenesis in cell culture. Later, cells-fibrin constructs were made and implanted subcutaneously into nude mice for 5 wks, then explanted for histological examination and glycosaminoglycans (GAG) quantification. Test group constructs  showed  higher  cartilage  matrix synthesis  as confirmed by Safranin O staining and GAG production. These results demonstrated the effectiveness of low serum media supplemented with TGF-ß3 in promoting  chondrogenesis in BMSCs.