Investigating On Characterization Of Golden Monkey Dermal Fibroblasts Of The Ear And Induction Of It Into The Germ Cells

Snub nosed monkey is listed as one of the most important protected animals in China, and it is also the second national treasure of China. Due to the small number of it, the state promulgated the relevant laws and regulations such as "Forest law" and the "Wildlife Protection Ordinance", and it has made the related publicity and education work, but the monkey survival pressure still can not be ignored. Most method of the protection of Sichuan golden monkey research are ecology macro methods, but the perspective of on cell biology research is rare. This experiment aims to induce Sichuan golden monkey dermal fibroblasts into germ lineage line cells with the method of molecular induction and tissue extract induction.(1) The cultivation,separation and identification of the Sichuan golden monkey dermal fibroblasts. The results showed that the morphology of the cells was consistent with fibroblasts. After cell generation, the Sichuan golden monkey dermal fibroblasts had good cell viability. Using PCR and flow cytometry technology tested the fibroblasts and mesenchymal cell surface marker protein, Marker gene prrxl, twist2, ddr2, runx2 are positive expression.The results showed that the expression of CD29 and CD90 was high, and the expression of CD79 was not expressed in the dermal fibroblast, It is in accordance with fibroblast cell types. The results showed that the cultured cells were fibroblasts.(2) Induce the cells by Tranylcypromine?FSK?repsox?VPA?AM580?EP2004777?Chir99021, It was found that the cells showed a vortex like shape, and some of the cells changed from spindle to round. PCR and qPCR detection of marker genes sall4 and gata6, It is indicated that PCR can run out the marker gene,qPCR result showed from 6th to 8th day,The expression level of gata6 was significantly higher than that of untreated control group(p<0.05), the expression level of sall4 was significantly higher than that of control group(p<0.01). It has been proved that the cells have been induced to differentiation towards the endoderm lineage cells.(3) The cells were treated by small molecules then induced of mouse testis tissue with 57 days, During this period, We did observation and gene detection of the cells. The results showed that the cells in the tissue extract were induced to 15 days, The levels of vasa and sall4 were significantly higher than those of control group (p<0.05),The endoderm lineage cells by the first stage have been induced to differentiation towards germ lineage cells after the induction of testicular tissue.In summary, In this study,we successfully isolated and cultured the golden monkey fibroblasts,After the induction of small molecular and tissue extracts.It is proved that the first stage of the small molecule induction can successfully induce the dermal fibroblasts of Sichuan golden monkey differentiate towards the endoderm lineage cells, and it was suggested that the second stage induction of mouse testicular tissue extracts could induce the endoderm lineage cells differentiate towards germ lineage cells.