Comparison Of The Effects Of Different Culture Systems On The Proliferation Of Chicken Spermatogonial Stem Cells In Vitro
Keywords
Collagen, Fibronectin, Laminin, Sertoli Cells, Chicken, Spermatogonial Stem Cells
Abstract
This study compared the effects of laminin, fibronectin, collagen and sertolicell on the in vitro proliferation of chicken (gallusgallus) spermatogonial stem cells (sscs) in four different culture systems. effect. SSCs were isolated using a three-enzyme two-step digestion method. The cells were cultured after differential purification with gelatin. The SSCs passed to the third generation were re-seeded into laminin, fibronectin and collagen-coated culture dishes and testicular Sertoli cells were prepared. On the feeder layer, the effects of different culture systems on the in vitro proliferation of gallinogen spermatozoa stem cells were tested through morphology, 5-ethyl-2′-deoxyuridine (edu) cell proliferation, and qrt-PCR technology. The results showed that the number of alkaline phosphatase (AKP)-positive clones in chicken SSCs was the highest in the testicular Sertoli cell group, reaching (32¡À3) per visual field, and the laminin, fibronectin, and collagen groups were (26) respectively. ¡À3), (24¡À2) and (23¡À2), the difference between the three groups was not significant (p>0.05); the cell proliferation rate of the testicular Sertoli cell group measured by edu was (92.82¡À2.15)%, which was significantly higher than the three groups Matrix proteome (p<0.01); qrt-PCR results showed that the proliferation marker genes proto-oncogene (myconcogene, c-myc), kruppel-like factor 4 (klf4), and proliferating cell nuclear antigen (pcna) were The testicular Sertoli cell group has the highest expression, followed by the laminin group. The expression of c-myc and klf4 in the fibronectin group is higher than that in the collagen group, and the opposite is true for pcna. The experimental results showed that all three groups of matrix proteins and testicular Sertoli cells can promote the proliferation of chicken SSCs in vitro, among which testicular Sertoli cells have the best effect, followed by laminin, fibronectin and collagen. The difference in effect is not significant. This result provides experimental basis and theoretical support for further optimizing the in vitro culture system of chicken embryo SSCs and elucidating the mechanism of germ cell proliferation. For further information of this article and research, feel free to contact our team for asssitance. Original research was done by Shi Qingqing, Zuo Qisheng, Li Dong, Zhang Lei, Huang Xiaomei, Zhang Zhentao, Zhang Yani, Li Bichun
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