1.Effect of brain microvascular endothelial cells on differentiation of co-cultured mesenchymal stem cells under hypoxic conditions
Kaiming LIU ; Luxiang CHI ; Xianghui LU
Journal of Third Military Medical University 2003;0(07):-
Objective To investigate the effect of brain microvascular endothelial cells (BMECs) on mesenchymal stem cells (MSCs) expressing fetal liver kinase-1(Flk-1) and von Willebrand factor (vWF) under normal and hypoxic conditions. Methods Human bone marrow MSCs and rat BMECs were isolated, cultured and identified. We used direct and indirect co-culture to induce the differentiation of MSCs. The expression of Flk-1 and vWF after induction was observed and analyzed by flow cytometry and immunofluorescence staining. Results Undifferentiated MSCs did not express Flk-1 or vWF. In indirect co-culture group, indirect co-culture under hypoxic conditions induced (7.58?0.58)% (n=6, P=0.034) MSCs to express Flk-1 but not vWF, while in normoxic indirect co-culture, MSCs expressed neither. In direct co-culture group, after 5-day direct co-culture with BMECs under normoxia or hypoxia, MSCs expressing Flk-1 accounted for (13.76?1.67)% (n=6, P
2.Paracrine actions of mesenchymal stem cells to the function of brain microvascular endothelial cells in hypoxic conditions
Kaiming LIU ; Luxiang CHI ; Xianghui LU
Chinese Journal of Pathophysiology 2000;0(07):-
AIM:To investigate the paracrine actions of mesenchymal stem cells(MSCs)to brain microvascular endothelial cells(BMECs)in hypoxic conditions.METHODS:Human bone marrow MSCs and rat BMECs were isolated,cultured and identified.The contents of VEGF and MMP-9 were confirmed using ELISA assays of the conditioned media.The transendothelial electrical resistance(TEER)was detected to reflect the permeability of the BMECs monolayer.The effects of different conditioned media on BMECs were investigated in hypoxic conditions.RESULTS:The contents of VEGF and MMP-9 in conditioned media were induced to increase significantly by the hypoxic stress,which were much lower in BMECs conditioned media than that in MSCs conditioned media.Hypoxia induced a significant increase in VEGF and MMP-9 in MSCs conditioned media collected in comparison to those in normal conditions.In hypoxic conditions,MSCs conditioned media enhanced the proliferation(optical density values:0.947?0.103 vs 0.254?0.024,P
3.Apoptosis in Adult Mouse Brain after Chronic Poisoning of Ketamine
Ju YANG ; Xiaojing LI ; Zhixiang ZHANG ; Kaiming LU ; Shizhong BIAN
Journal of Forensic Medicine 2013;(5):325-329
Objective To study the effect of chronic poisoning of ketamine on brain cell apoptosis in adult mouse under different duration and doses. Methods The mouse model of chronic poisoning of ketamine was established on adult mouse by tail vein injection of ketamine twice every week with different doses (4, 10, 20 and 30 m g/kg). The mice were sacrificed after continuous injection of ketamine of 1, 2, 4, 8 and 12 weeks. The qualitative assessment of apoptosis was made by transmission electron microscope and the quantitative assessment was made by Caspase-3 im m umofluorescence staining method and terminal deoxynucleotidyl transferase-mediated dU TP nick end labeling (TUNEL ) to estimate the time point of apoptosis. All the experimental results were statistically analyzed. Results The neuron apoptosis was ob-served in hippocam pus and corpus striatum by transmission electron microscope one week after adminis-tration, and continued for eight weeks. High level of Caspase-3 expression was observed one week after administration, but with a lowlevel expression after 4 weeks. The num ber of TUNEL positive cells ob-viously increased one week after administration and maintained in ahigh num ber at 4 weeks. Conclu-sion Ketamine by tail vein injection could induce neuron apoptosis in adult mouse.