[Objective] The aim was to explore the effect of cumulus cells on the in vitro fertilization of in vitro matured bovine oocytes. [Method] The in vitro matured oocytes were divided into three groups of cumulus cells re...[Objective] The aim was to explore the effect of cumulus cells on the in vitro fertilization of in vitro matured bovine oocytes. [Method] The in vitro matured oocytes were divided into three groups of cumulus cells removal, partial removal and no removal. [Result] In the co-culture with cumulus cells, the oocytes of the removal group had higher cleavage rate and blastocyst rate （74.4%±4.1, 53.7%±5.1） than those of the no removal group （72.7%±5.1, 52.4%±3.5）, but the difference was not significant （P〉0.05）, while both groups had better performances than the re- moval group （39.6%±4.5, 18.8%±4.6） with the difference reaching the significant level （P〈0.05）. All the three groups showed significant difference with the control. The combination of cumulus cells and melatonin achieved the best effects as the cleavage rate and blastocyst rate of the partial removal group （79.8%±3.7, 56.5%±5.1） were better than those of the no removal group （78.2%±2.6, 55.8%±4.6）, and the difference was not significant, while both group had better performances than the removal group （48.3%±5.5, 22.7%±4.3） and the control group with the differences reaching the significant level （P〈0.05）. [Conclusion] The study provided technical support for the production of dairy cows and beef cattle.展开更多
To prepare a hand-made micropore membrane culture plate insert forco-culture. Methods The plate insert was made using plastic centrifuge tube and micropore membrane.After seeding brain capillary endothelial cells (BCE...To prepare a hand-made micropore membrane culture plate insert forco-culture. Methods The plate insert was made using plastic centrifuge tube and micropore membrane.After seeding brain capillary endothelial cells (BCECs) on it (under the effect ofastrocyte-conditioned medium), the plate insert was assessed by analysis of trans-endothelialelectrical resistance (TEER). Results The plate insert has a stability of at least 15 d underculture condition. TEER increased significantly under co-culture condition from (66.1 +- 13.3)Ωcm^2 to (182.2 +- 6.7) Ωcm^2. Conclusion This micropore membrane culture plate insert can beeasily made, on which BCEC culture can be successfully performed. Moreover, it is adjustable andrecyclable. It follows that the plate insert is a useful tool for co-culture and the relatedresearch fields.展开更多
Objective： By establishing the indirect contact co-culture system, we studied the in vitro condition for MAPCs differentiating into epidermal cells and the transformation of MAPCs into epidermal cell phenotype. Meth...Objective： By establishing the indirect contact co-culture system, we studied the in vitro condition for MAPCs differentiating into epidermal cells and the transformation of MAPCs into epidermal cell phenotype. Methods： Cell culture insert membrane was used for substitute basal membrane and MAPCs, fibroblast cells （FCs） and mixture of MAPCs and epidermal cells and FCs were separately implanted into 2 sides of it. PKH26 was used to label cloned MAPCs; type IV collagen rapid adhering method was used to isolate and culture the skin epidermal cells from l-day-old SD rat. Results： Part of the MAPCs transformed into cells expressing keratin in the presence of peripheral epithelia and FCs. Type Ⅳ collagen rapid adhering method successfully selected rats＇ epidermal stem cells. The mixture of the 2 kinds of cells or indirect culture might promote the differentiation through mesenchymal factors secreted by dermis FC. Conclusion： We were the first to have established the in vitro model of MAPCs differentiation into epidermal cells, in which MAPCs were transformed into epithelium-like cells.展开更多
The breakthrough development of induced pluripotent stem cells(iPSCs)raises the prospect of patient-specific treatment for many diseases through the replacement of affected cells.However,whether iPSC-derived functiona...The breakthrough development of induced pluripotent stem cells(iPSCs)raises the prospect of patient-specific treatment for many diseases through the replacement of affected cells.However,whether iPSC-derived functional cell lineages generate a deleterious immune response upon auto-transplantation remains unclear.In this study,we differentiated five human iPSC lines from skin fibroblasts and urine cells into neural progenitor cells(NPCs)and analyzed their immunogenicity.Through co-culture with autogenous peripheral blood mononuclear cells(PBMCs),we showed that both somatic cells and iPSC-derived NPCs do not stimulate significant autogenous PBMC proliferation.However,a significant immune reaction was detected when these cells were co-cultured with allogenous PBMCs.Furthermore,no significant expression of perforin or granzyme B was detected following stimulation of autogenous immune effector cells(CD3+CD8 T cells,CD3+CD8+T cells or CD3 CD56+NK cells)by NPCs in both PBMC and T cell co-culture systems.These results suggest that human iPSC-derived NPCs may not initiate an immune response in autogenous transplants,and thus set a base for further preclinical evaluation of human iPSCs.展开更多
基金Supported by the Key Program for Agriculture of Qiqihar City(NYGG-201524)~~
文摘[Objective] The aim was to explore the effect of cumulus cells on the in vitro fertilization of in vitro matured bovine oocytes. [Method] The in vitro matured oocytes were divided into three groups of cumulus cells removal, partial removal and no removal. [Result] In the co-culture with cumulus cells, the oocytes of the removal group had higher cleavage rate and blastocyst rate （74.4%±4.1, 53.7%±5.1） than those of the no removal group （72.7%±5.1, 52.4%±3.5）, but the difference was not significant （P〉0.05）, while both groups had better performances than the re- moval group （39.6%±4.5, 18.8%±4.6） with the difference reaching the significant level （P〈0.05）. All the three groups showed significant difference with the control. The combination of cumulus cells and melatonin achieved the best effects as the cleavage rate and blastocyst rate of the partial removal group （79.8%±3.7, 56.5%±5.1） were better than those of the no removal group （78.2%±2.6, 55.8%±4.6）, and the difference was not significant, while both group had better performances than the removal group （48.3%±5.5, 22.7%±4.3） and the control group with the differences reaching the significant level （P〈0.05）. [Conclusion] The study provided technical support for the production of dairy cows and beef cattle.
基金NationalMedicine 863Project (No .2 0 0 2AA2Z3 43C)
文摘To prepare a hand-made micropore membrane culture plate insert forco-culture. Methods The plate insert was made using plastic centrifuge tube and micropore membrane.After seeding brain capillary endothelial cells (BCECs) on it (under the effect ofastrocyte-conditioned medium), the plate insert was assessed by analysis of trans-endothelialelectrical resistance (TEER). Results The plate insert has a stability of at least 15 d underculture condition. TEER increased significantly under co-culture condition from (66.1 +- 13.3)Ωcm^2 to (182.2 +- 6.7) Ωcm^2. Conclusion This micropore membrane culture plate insert can beeasily made, on which BCEC culture can be successfully performed. Moreover, it is adjustable andrecyclable. It follows that the plate insert is a useful tool for co-culture and the relatedresearch fields.
基金Supported by the National Natural Science Foundation of China(30600651)the Cooperation Foundation for Overseas Young Scientists (30428001)
文摘Objective： By establishing the indirect contact co-culture system, we studied the in vitro condition for MAPCs differentiating into epidermal cells and the transformation of MAPCs into epidermal cell phenotype. Methods： Cell culture insert membrane was used for substitute basal membrane and MAPCs, fibroblast cells （FCs） and mixture of MAPCs and epidermal cells and FCs were separately implanted into 2 sides of it. PKH26 was used to label cloned MAPCs; type IV collagen rapid adhering method was used to isolate and culture the skin epidermal cells from l-day-old SD rat. Results： Part of the MAPCs transformed into cells expressing keratin in the presence of peripheral epithelia and FCs. Type Ⅳ collagen rapid adhering method successfully selected rats＇ epidermal stem cells. The mixture of the 2 kinds of cells or indirect culture might promote the differentiation through mesenchymal factors secreted by dermis FC. Conclusion： We were the first to have established the in vitro model of MAPCs differentiation into epidermal cells, in which MAPCs were transformed into epithelium-like cells.
基金supported by the National Basic Research Program of China,Ministry of Science and Technology(2011CB965204,2012CB966802)the National Natural Science Foundation of China(31000402)+3 种基金the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA01020401,XDA-01020202)the Ministry of Science and Technology International Technology Cooperation Program(2012DFH30050)the National Science&Technology Major Special Project on Major New Drug Innovation(2011ZX09102-010-01)the Development and Technology Innovation for Equipment Functional Development Project of Chinese Academy of Sciences(yg2011082,yg2011083)
文摘The breakthrough development of induced pluripotent stem cells(iPSCs)raises the prospect of patient-specific treatment for many diseases through the replacement of affected cells.However,whether iPSC-derived functional cell lineages generate a deleterious immune response upon auto-transplantation remains unclear.In this study,we differentiated five human iPSC lines from skin fibroblasts and urine cells into neural progenitor cells(NPCs)and analyzed their immunogenicity.Through co-culture with autogenous peripheral blood mononuclear cells(PBMCs),we showed that both somatic cells and iPSC-derived NPCs do not stimulate significant autogenous PBMC proliferation.However,a significant immune reaction was detected when these cells were co-cultured with allogenous PBMCs.Furthermore,no significant expression of perforin or granzyme B was detected following stimulation of autogenous immune effector cells(CD3+CD8 T cells,CD3+CD8+T cells or CD3 CD56+NK cells)by NPCs in both PBMC and T cell co-culture systems.These results suggest that human iPSC-derived NPCs may not initiate an immune response in autogenous transplants,and thus set a base for further preclinical evaluation of human iPSCs.
