Comparison between Single and Co-culture of Adipocyte and Muscle Cell Lines in Cell Morphology and Cytosolic Substances
2012
Choi, C.W., Daegu University, Gyeongsan, Republic of Korea | Cho, W.M., National Institute of Animal Science, RDA, Suwon, Republic of Korea | Yeon, S.H., National Institute of Animal Science, RDA, Suwon, Republic of Korea | Hwangbo, S., Daegu University, Gyeongsan, Republic of Korea | Song, M.K., Chungbuk National University, Cheongju, Republic of Korea | Park, S.K., National Institute of Animal Science, RDA, Suwon, Republic of Korea | Baek, K.H., Yeungnam University, Gyeongsan, Republic of Korea
Present study was performed to investigate the effect of single and co-culture of adipocyte and muscle cell lines on cell differentiation. 3T3-L1 (adipocyte) and L6 (muscle) cell lines were single-cultured on the condition of 10% fetal bovine serum (FBS)/Dulbeco's modified eagle's medium (DMEM) for 48 h followed by culture within 5% FBS/DMEM as a growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without additives in single- or co-culture of the 3T3-L1 and the L6 cells to induce differentiation of both cell types. In co-culture system, the 3T3-L1 and the L6 cells were grown in separated places by being seeded on a 0.4 ㎛ insert membrane and on the bottom of 6 well plate, respectively. Cell differentiation was measured using morphological investigation and cytosolic analysis of glycerol-3-phosphate dehydrogenase (GPDH; for 3T3-L1) and creatine kinase (CK; for L6). Based on the GPDH results, the presence of L6 cells did not stimulate 3T3-L1 differentiation showing more differentiation of 3T3-L1 cells in the single-culture compared to the co-culture condition. In contrast, 3T3-L1 cells in the co-culture promoted differentiation of L6 cells. Enzymatic analysis supported this result showing that 3T3-L1 cells showed statistically (P less than 0.05) higher GPDH activity in the single-culture than the co-culture, whereas CK results of L6 cells were vice versa (P less than 0.05). Overall, present results may indicate that co-culture system is more reliable and precise technique compared to single-culture. Further studies on several co-culture trials including different media conditions, supplementation of differentiating substances, molecular biological analysis, etc. should be required to obtain practical and fundamental mass data.
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