Effect of Culture Condition on Chondrocyte Viability Isolated from Articular Cartilage

Kim, Sang Gyung; Choe, Jung Yoon; Choi, Chang Hyuk; Choi, Gi Hwan; Kim, Jong Ki

Abstract

BACKGROUND
Recent advance in tissue engineering in the biomedical field shed light on the replacement or regeneration of various organs with synthetic substitutes. Currently emerging cartilage tissue engineering therapies involve artificial cartilage fabricated from three dimensional cultures using appro-priate scaffolds. It is mandatory to expand or proliferate the chondrocytes in vitro to prepare the artificial cartilage. The purpose of this study was to find out the most favorable culture conditions for chon-drocyte viability in vitro. METHODS: Articulr chondrocytes or cartilage explants were isolated from the patellofemoral groove of adult pigs. And then we standardized the size and thickness of the cartilage explants as well as preparing alginate-chondrocyte beads for three-dimensional cultures. The cartilage explants, including 10% fetal bovine serum for 10 days, 36 days and passage 6. Cellualr viability was measured by methylthiazol tetrazolium (MTT) assay on monolayer, alginate bead and cartilage explant. SPSS 11.5 was used for data anaylsis. RESULTS: Chondrocytes cultured on monolayers in vitro showed no significant difference in cellular viability until passage 6 following isolation from the patellofemoral groove of adult pigs (P>0.05, n=4). Chondrocyte viability was markedly increased by day 16 both in the monolayer (148%) and three dimensional cultures (245%), and then slightly decreased 126% and 200%, respectively, at day 36. Three dimensional cultures using alginate bead were more favorable for chodrocyte viability than monolayer culture in chondrocyte primary culture (P=0.003, n=6). Chondrocyte viability in the algi-nate bead was increased 300% during 36 days' incubation period (P=0.001, n=3). Cellular viability in the cartilage explant culture was decreased after day 4 in both MTT score (P=0.022, n=10) and MTT OD (P=0.039, n=10). CONCLUSIONS: Three dimensional cultures using alginate bead were the most favorable for chon-drocyte viability in chondrocyte primary cultures.