Blood samples were collected from chicken embryos at stage 11～15, and labeled with fluorescent dye PKH26. Primordial germ cells (PGCs) were then isolated from blood samples by nycodenz density gradient centrifugation...Blood samples were collected from chicken embryos at stage 11～15, and labeled with fluorescent dye PKH26. Primordial germ cells (PGCs) were then isolated from blood samples by nycodenz density gradient centrifugation. After PGCs were labeled and isolated, about 200 PGCs in one microliter were injected into the subgerminal cavity of quail blastoderm at stageⅩ. After 48 hours incubation, chicken PGCs were identified by fluorescent microscopy. Red fluorescence emitted from PKH26 labeled chicken PGCs was observed in the head, the heart and the developing gonadal anlage of quail embryos. The result suggests that chicken PGCs still keep migration ability after 56 hours.展开更多
Primordial germ cells(PGCs) isolated from gonads of stage 19 by Ficoll density gradient centrifugation were cultured in DMEM medium in vitro.Then the second passage PGCs were induced to differentiate into neuron-like ...Primordial germ cells(PGCs) isolated from gonads of stage 19 by Ficoll density gradient centrifugation were cultured in DMEM medium in vitro.Then the second passage PGCs were induced to differentiate into neuron-like cells by different induction media.Specific markers and structures were detected by histochemistry method.The results showed that induction medium RA and IBMX were the best ways to induce PGCs into neuron-like cells,and almost 80%-85% cells exhibited typical neuron-like phenotype after induction.Special Nissl body was found by histochemistry.The effect of induction medium RA+IBMS,RA+BHA+DMSO and RA+BHA+DMSO+IBMX were better than that of induction medium BHA+DMSO.Under the same induction medium,about 80%-85% neuron-like cells could be detected no matter what PGCs were cultured 24 h,48h or 72 h in vitro.展开更多
文摘Blood samples were collected from chicken embryos at stage 11～15, and labeled with fluorescent dye PKH26. Primordial germ cells (PGCs) were then isolated from blood samples by nycodenz density gradient centrifugation. After PGCs were labeled and isolated, about 200 PGCs in one microliter were injected into the subgerminal cavity of quail blastoderm at stageⅩ. After 48 hours incubation, chicken PGCs were identified by fluorescent microscopy. Red fluorescence emitted from PKH26 labeled chicken PGCs was observed in the head, the heart and the developing gonadal anlage of quail embryos. The result suggests that chicken PGCs still keep migration ability after 56 hours.
文摘Primordial germ cells(PGCs) isolated from gonads of stage 19 by Ficoll density gradient centrifugation were cultured in DMEM medium in vitro.Then the second passage PGCs were induced to differentiate into neuron-like cells by different induction media.Specific markers and structures were detected by histochemistry method.The results showed that induction medium RA and IBMX were the best ways to induce PGCs into neuron-like cells,and almost 80%-85% cells exhibited typical neuron-like phenotype after induction.Special Nissl body was found by histochemistry.The effect of induction medium RA+IBMS,RA+BHA+DMSO and RA+BHA+DMSO+IBMX were better than that of induction medium BHA+DMSO.Under the same induction medium,about 80%-85% neuron-like cells could be detected no matter what PGCs were cultured 24 h,48h or 72 h in vitro.
