Ngoc Tran T D, Stovall K E, Suantawee T, Hu Y, Yao S, Yang L-J, Adisakwattana S, Cheng H
Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA, USA.
Program in Biomedical Sciences, Graduate School, Chulalongkorn University, Bangkok, Thailand.
Cell Prolif. 2017 Oct;50(5). doi: 10.1111/cpr.12360. Epub 2017 Jul 30.
Investigate the role of the transient receptor potential melastatin 4 (TRPM4) channel in rat dental pulp stem cell (DPSC) proliferation and survival.
Immunofluorescence and FACS analysis were used to detect the stem cell marker CD90. Alizarin Red S and Oil Red O staining were used to identify osteoblast and adipocyte differentiation, respectively. To characterize TRPM4, patch-clamp recordings were obtained from single cells in the whole-cell configuration mode. The significance of TRPM4 for proliferation and survival was examined with 9-phenanthrol, a TRPM4 inhibitor during a 96-hour period of culture. Real-time Ca imaging analysis with Fura-2AM was used to investigate the impact of TRPM4 on intracellular Ca signals.
DPSCs were CD90-positive and differentiated into osteoblasts. Patch-clamp recordings revealed currents typical of TRPM4 that were Ca -activated, voltage-dependent and Na -conducting. Inhibition of TRPM4 resulted in a significant reduction in the cell population after a 96-hr period of culture and transformed the biphasic pattern of intracellular Ca signalling into sustained oscillations.
Rat DPSCs have stem cell characteristics and functional TRPM4 channels that are required for proliferation and survival. These data suggest that the shape and frequency of intracellular Ca signals may mediate stem cell proliferation and survival.
研究瞬时受体电位褪黑素4(TRPM4)通道在大鼠牙髓干细胞(DPSC)增殖和存活中的作用。
采用免疫荧光和流式细胞术分析检测干细胞标志物CD90。分别用茜素红S和油红O染色鉴定成骨细胞和脂肪细胞分化。为了表征TRPM4,采用全细胞模式从单细胞获得膜片钳记录。在96小时的培养期间,用TRPM4抑制剂菲咯啉研究TRPM4对增殖和存活的意义。使用Fura-2AM进行实时钙成像分析,以研究TRPM4对细胞内钙信号的影响。
DPSCs呈CD90阳性并分化为成骨细胞。膜片钳记录显示出典型的TRPM4电流,其为钙激活、电压依赖性且对钠有通透性。在96小时的培养期后,抑制TRPM4导致细胞数量显著减少,并将细胞内钙信号的双相模式转变为持续振荡。
大鼠DPSCs具有干细胞特性和功能性TRPM4通道,这些通道是增殖和存活所必需的。这些数据表明,细胞内钙信号的形状和频率可能介导干细胞的增殖和存活。