Summary： To isolate and culture the purified monoclonal neural stem cells from the cerebral cortex of new born mice, new-born mice cerebral cortex was isolated and dissociated to single-cell suspension by mechanical ...Summary： To isolate and culture the purified monoclonal neural stem cells from the cerebral cortex of new born mice, new-born mice cerebral cortex was isolated and dissociated to single-cell suspension by mechanical trituration. The dissociated single cells were cultured in serum-free medium. After the formation of neurospheres, single-cell clone culture was performed by limiting dilution and the proliferated single-cell clones were harvested for subculture. Immunocytochemistry was used to detect the specific marker of neuroepithelial stem cells （Nestin） of the primary and monoclonal neurospheres. In the differentiated cells we detected the specific antigen of NF-200 and GFAP. Our results showed that the primary neurospheres expressed Nestin antigen positively. By limiting dilution, we cultured the cell lines from single-cell clone and the monoclonal neurospheres expressed Nestin and had capabilities of self-renewal, proliferation and the potentiality of differentiation into neurons and glial cells. It is concluded that monoclonal neural stem cells which have the ability of proliferation and multi-directional differentiation can be isolated and cultured from the cerebral cortex of new-born mice by limiting dilution.展开更多
Nannochloropsis is a genus of marine eukaryotic unicellular algae,which belongs to class Eustigmatophyceae.The spe-cies of Nannochloropsis which are fine rotifer feed and rich in eicosapentaenoic acid(EPA)are economic...Nannochloropsis is a genus of marine eukaryotic unicellular algae,which belongs to class Eustigmatophyceae.The spe-cies of Nannochloropsis which are fine rotifer feed and rich in eicosapentaenoic acid(EPA)are economically important.Species in this genus are usually 2-5μm in size and are morphologically similar,which makes their identification difficult.We obtained a monoclone of Nannochloropsis with plating method in this study.DNA was extracted and the quality was determined by restriction enzyme digestion and spectrophotometer analysis.The DNA extracted was used to amplify the sequences of 18S ribosomal RNA gene,ITS region of ribosomal RNA transcription unit and rbcL gene.The phylogenetic analysis was carried out by constructing the neighbor-joining trees with Tamura-Nei distances.The phylogenetic analysis showed that the monoclone is N.oceanica.展开更多
Preliminary study has been made to test wether three strains McAb（1B1,5D<sub>6</sub>, 6D<sub>8</sub>）are against the same antigen determinant.Througn ELISA additive and competition expeiments...Preliminary study has been made to test wether three strains McAb（1B1,5D<sub>6</sub>, 6D<sub>8</sub>）are against the same antigen determinant.Througn ELISA additive and competition expeiments,itproved that these three strains are against different antigen determinant.The result of positiveserum antigen component analysis with 2 strains IBDV McAb showed that sample IBD positiveserum had obvious inhibition against combination of 5D<sub>6</sub> McAb with corresponding antigen.Theresults of substitution of corresponding component in ELISA inhibition experiment and compari-son of non-IBD serum （SPF chicken serum,ND,MD,IA positive serum）proved that IBD anti-serum was the only one showing inhibition against 5D<sub>6</sub> McAb.Comparison with AGP and electro-microscope observation showed that ELISA inhibition experiment was characterised by high-specificity,rapidity and sensitiveness.799 serum samples were tested with ELISA inhibition anddouble immunodiffusion（AGP） experiments.ELISA had gotten 486 positive,with positive rate of60.83%;and AGP 334。展开更多
基金This project was supported by a grant from the National Natural Science Foundation of China (No.30371397)
文摘Summary： To isolate and culture the purified monoclonal neural stem cells from the cerebral cortex of new born mice, new-born mice cerebral cortex was isolated and dissociated to single-cell suspension by mechanical trituration. The dissociated single cells were cultured in serum-free medium. After the formation of neurospheres, single-cell clone culture was performed by limiting dilution and the proliferated single-cell clones were harvested for subculture. Immunocytochemistry was used to detect the specific marker of neuroepithelial stem cells （Nestin） of the primary and monoclonal neurospheres. In the differentiated cells we detected the specific antigen of NF-200 and GFAP. Our results showed that the primary neurospheres expressed Nestin antigen positively. By limiting dilution, we cultured the cell lines from single-cell clone and the monoclonal neurospheres expressed Nestin and had capabilities of self-renewal, proliferation and the potentiality of differentiation into neurons and glial cells. It is concluded that monoclonal neural stem cells which have the ability of proliferation and multi-directional differentiation can be isolated and cultured from the cerebral cortex of new-born mice by limiting dilution.
基金the National High Technology Research and Development Program of China (2010AA10A403)the National Natural Science Foundation of China (No.40976076)+1 种基金the Major State Basic Research Development Program of China (973 Program) (2011CB200901)the Basic Research Program of Municipal Bureau of Science and Technology of Qingdao (09-1-3-22-jch)
文摘Nannochloropsis is a genus of marine eukaryotic unicellular algae,which belongs to class Eustigmatophyceae.The spe-cies of Nannochloropsis which are fine rotifer feed and rich in eicosapentaenoic acid(EPA)are economically important.Species in this genus are usually 2-5μm in size and are morphologically similar,which makes their identification difficult.We obtained a monoclone of Nannochloropsis with plating method in this study.DNA was extracted and the quality was determined by restriction enzyme digestion and spectrophotometer analysis.The DNA extracted was used to amplify the sequences of 18S ribosomal RNA gene,ITS region of ribosomal RNA transcription unit and rbcL gene.The phylogenetic analysis was carried out by constructing the neighbor-joining trees with Tamura-Nei distances.The phylogenetic analysis showed that the monoclone is N.oceanica.
文摘Preliminary study has been made to test wether three strains McAb（1B1,5D<sub>6</sub>, 6D<sub>8</sub>）are against the same antigen determinant.Througn ELISA additive and competition expeiments,itproved that these three strains are against different antigen determinant.The result of positiveserum antigen component analysis with 2 strains IBDV McAb showed that sample IBD positiveserum had obvious inhibition against combination of 5D<sub>6</sub> McAb with corresponding antigen.Theresults of substitution of corresponding component in ELISA inhibition experiment and compari-son of non-IBD serum （SPF chicken serum,ND,MD,IA positive serum）proved that IBD anti-serum was the only one showing inhibition against 5D<sub>6</sub> McAb.Comparison with AGP and electro-microscope observation showed that ELISA inhibition experiment was characterised by high-specificity,rapidity and sensitiveness.799 serum samples were tested with ELISA inhibition anddouble immunodiffusion（AGP） experiments.ELISA had gotten 486 positive,with positive rate of60.83%;and AGP 334。
文摘目的为抗原抗体反应提供原子力显微镜下的形态学基础,为建立常用抗体形态结构的形态库打下基础。方法将7种常用单克隆抗体分别滴加于载玻片上,放入加有10%中性缓冲甲醛的湿盒内,置于37℃温箱中1 h,用蒸馏水轻轻涮洗,晾干,用原子力显微镜(atomic force microscope,AFM)进行扫描,观察形态,测量扫描颗粒的宽度、最大峰高度和平均粗糙度。结果在2D图中,细胞角蛋白(广谱)、S100蛋白鼠抗人单克隆抗体为长杆状,波形蛋白、白细胞共同抗原、细胞角蛋白7、Dog-1鼠抗人单克隆抗体为类圆形,甲状腺球蛋白鼠抗人单克隆抗体为三角形;不同组间单克隆抗体之间宽度、最大峰高度的差异有统计学意义(P<0.05),各组间平均粗糙度的差异无统计学意义(P>0.05)。结论 2D图中,7种单克隆抗体的形态大致可分为三种:长杆状、类圆形、三角形,直观地印证了抗体具有相对特异性,并为抗原的共同表达和例外表达的进一步研究提供形态学基础。
