ABSTRACT
Objective The changes of Mg2+ concentration and cell apoptosis were detected by using siRNA to silence MagT1 in rat cardio myocytes.Methods MagT1 siRNA sequences were designed and synthetized,then transfected into primary cultured rat myocardial cell for silencing MagT1.The expression of MagT1 mRNA and protein were detected by RT-PCR and Western blot.The changes of Mg2+ concentration in the cells were detected by fluorescence microscopy.Flow cytometry (FCM) was used to detect the cell apoptosis.Results Compared with negative siRNA group,MagT1 siRNA transfected rat cardiomyocytes after 48 h,MagT1 mRNA silence efficiency was 51.83 ％ (P＜0.05),the silence of MagT1 protein efficiency was 56.75 ％ (P＜0.05),intracellular Mg2+ concentration was reduced by 29.13％ (P＜0.05),the apoptosis rate was 31.18％ (P＜0.01);MagT1 siRNA transfected rat cardiomyocytes after 60 h,MagT1 mRNA silence efficiency was 86.91％ (P＜0.01),the silence of MagT1 protein efficiency was 83.85％ (P＜0.01),intracellular Mg2+ concentration was reduced by 41.32％ (P＜0.01),the apoptosis rate was 40.61％ (P＜0.01).Conclusion After the silencing of MagT1,the concentration of Mg2+ in the cells decreased significantly,the apoptotic rate increased significantly,cell life activities are greatly affected.
ABSTRACT
Objective To establish a culture system in vitro of adult human retinal neural cells for providing a model for the research of retinal neural cells. Design Experimental study. Participants Cultured adult human retinal neural cells. Methods The isolated cells from adult human postmortem retina (20?40 years old) were cultured, then cells of different stages were identified with immunocytochemical staining and judged with phase contrast microscopy and electron microscopy. Main Outcome Measures Cellular morphology and structure. Results (1) The results of cell culture: the adult retinal neural cells could survive in vitro under some conditions and were identified as NSE positive mostly. (2) The results of electron microscopy: most cultured cells were photoreceptors, bipolar cells, horizontal cells and some were glial cells with scanning electron microscopy. Conclusions Under feasible conditions, the adult human retinal neural cells could be cultured and maintained effectively in vitro.