1.Role of endoplasmic reticulum stress in ketamine-induced apoptosis in rat neurons: an in vitro experiment
Huan CHEN ; Di WEN ; Chunling MA ; Guanjie XU ; Qiongmei GUO ; Changhao ZHOU
Chinese Journal of Anesthesiology 2016;36(10):1225-1228
Objective To evaluate the role of endoplasmic reticulum stress in ketamine-induced apoptosis in rat neurons.Methods Rat adrenal pheochromocytoma cell line (PC12 cells) was seeded in the culture dishes 100 mm in diameter (10 ml/dish) or in 6-well plates (2 ml/well) at a density of 5 × 105 cells/ml.PC12 cells were divided into 4 groups (n =6 each) using a random number table:control group (group C);ketamine group (group K);endoplasmic reticulum stress inhibitor salubrinal group (group S);ketamine + salubrinal group (group K+S).In group C,the cells were cultured in the plain culture medium.In group K,1.5 mmol/L ketamine was added.In group S,30 mmol/L salubrinal was added.In group K + S,1.5 mmol/L ketamine and 30 mmol/L salubrinal were added.At 24 h of incubation,the cell morphology was observed under light microscope,the expression of Bip and caspase-12 in PC12 cells was detected by Western blot,and the cell apoptosis was measured by flow cytometry.The apoptosis rate was calculated.Results Compared with group C,the expression of Bip and caspase-12 was significantly upregulated,and the apoptosis rate was increased in K and K + S groups (P < 0.05),and no significant change was found in the parameters mentioned above in group S (P> 0.05).Compared with group K,the expression of Bip and caspase-12 was significantly down-regulated,and the apoptosis rate was decreased in group K+S (P<0.05).The degree of damage to PC12 cells was more serious in group K than in group C..The degree of damage to PC12 cells in group K+S was significantly mnilder than that in group K and more serious than that in group C.Conclusion The mechanism by which ketamine induces neuronal apoptosis is related to the enhancement of endoplasmic reticulum stress in rats.
2.Comparative study of human fetal and adult bone marrow derived mesenchymal stem cells.
Ying HU ; Li MA ; Guanjie MA ; Xueying JIANG ; Chunhua ZHAO
Chinese Journal of Hematology 2002;23(12):645-648
OBJECTIVETo explore the differences of phenotype and biological characteristics between fetal and adult bone marrow derived mesenchymal stem cells.
METHODSMononuclear cells from 4-5 months old human aborted fetus and normal adult bone marrow were cultured in SF medium to obtain mesenchymal stem cells. The growth curve, cell cycle, immunophenotype, in vitro expansion potential, differentiation capacities were investigated.
RESULTSThe adherent fetal and adult bone marrow-derived cells cultured in the absence of differentiation stimuli gave rise to a population of cells with phenotypical features of mesenchymal stem cells (MSC). These MSCs were similar in cell morphology and antigenic phenotype. The proliferative and multilineage differentiation potential of the bone marrow derived MSC from the fetus is higher than that from the adult, but the adherent ability of the MSCs from the adult is higher than that from the fetus.
CONCLUSIONFetal bone marrow derived MSCs should be enough to sustain a steady supply of low differentiated cells for proliferation, hence an abundant and accessible cellular reservoir for stem cell bioengineering, whereas adult bone marrow derived MSCs are more useful in hematopoietic reconstitution in bone marrow transplantation.
Adult ; Adult Stem Cells ; cytology ; immunology ; ultrastructure ; Bone Marrow Cells ; cytology ; immunology ; ultrastructure ; Cell Cycle ; Cell Differentiation ; Cell Lineage ; Cell Proliferation ; Cells, Cultured ; Fetal Stem Cells ; cytology ; immunology ; ultrastructure ; Flow Cytometry ; Humans ; Immunophenotyping ; Mesenchymal Stromal Cells ; cytology ; immunology ; ultrastructure ; Microscopy, Electron