Comparison of in vitro chondrogenic ability between two culture modes:pellet culture system and fibrin gel scaffold

HUANG Liang-jie; WENG Tu-jun; ZHANG Chun-li; LIU Yan; QIAN Long; HOU Shu-xun

doi:10.3969/j.issn.2095-252X.2017.03.008

Abstract：Objective To compare the effects of 2 different culture modes: pellet culture system and fibrin gel scaffold on the chondrocytic differentiation of adipose-derived stem cells ( ADSCs ) and the synthesis of cartilage extracellular matrix.Methods Adipose tissues were used to isolate human ADSCs and grown in Dulbecco's modified Eagle's medium ( DMEM ) containing 10% fetal bovine serum ( FBS ). After the human ADSCs were cultured to the 3rd generation, they were induced into cartilage by 2 kinds of culture modes respectively. At the 21st day, the cells were observed by hematoxylin-eosin ( HE ) staining and safranine-O staining. The glycosaminoglycan ( GAG ) content in extracellular matrix was determined by content dimeth-ylmethylene blue ( DMMB ) method. The expression of type Ⅱ collagen gene was assessed by real-time polymerase chain reaction ( PCR ).Results HE staining and safranine-O staining showed that the extracellular matrix of chondrocytes was strongly secreted in the fibrin gel scaffold group with transforming growth factor β3 ( TGFβ3 ) and fused into a sheet. It's indicated that the fibrin gel scaffold could better promote cartilage differentiation of stem cells and maintain chondrocyte type. The results of GAG / deoxyribonucleic acid ( DNA ) showed that the ability to secrete GAG in the fibrin gel scaffold group with TGFβ3 was better than that of the other group (P < 0.05 ), and the expression of type Ⅱ collagen messenger ribonucleic acid ( mRNA ) was the strongest (P < 0.05 ).Conclusions The ability of fibrin gel scaffold to differentiate ADSCs into cartilage is better than that of scaffold-free pellet culture system.