Effects of mycoplasma elimination agent on cardiac differentiation of human embryonic stem cells

Abstract

Mycoplasma contamination is a major problem in continuous cell culture. Various agents are used for the elimination of mycoplasma. However, their effects on cardiac differentiation potential of human embryonic stem cells (hESCs) are rarely known. In this study, we tried to evaluate the effects of mycoplasma elimination agent (plasmocin) on differentiation into cardiomyocytes (CMs) from hESCs. Undifferentiated hESCs were induced to differentiated into CMs in the presence of P/S and plasmocin (2.5, 5.0, 10.0 mg/ml). The expression of cardiac specific genes such as Nkx2.5, Tbx5, a-myosin heavy chain (MHC) and cardiac troponin (cTn) I was evaluated using immunocytochemistry and qPCR. After 14 days of differentiation, the beating cluster formation of hESCs was reduced in plasmocin-treated group. The expression of cardiac genes was down-regulated and the decrease of cTn I was significant in plasmocin-treated group. In this study, we demonstrated the effects of mycoplasma elimination agent on cardiac differentiation potential of hESCs. There results showed that addition of mycoplasma elimination agent reduced the differentiation potential of hESCs (Nos. 2010-0016819 and SC1150).