猪精原干细胞体外分离培养及诱导分化的研究

doi:10.11843/j.issn.0366-6964.2014.07.011

Abstract

Abstract:

The aim of this study was to establish the culture system of pSSC in vitro，and explore the expression of these key genes during the induced differentiation.Using the methods of differential adherent，we had isolated pSSCs from the supporting(Sertoli) cells.In addition，we also had identified the stem characteristics by the biochemical and immunological methods.When it grew to generation 3，the SSCs had been induced into the differentiation of adipocytes，neuron-like cells and osteoblast，through the addition of different chemical reagents；moreover，the results of induction also had been detected by the biochemical staining，quantitative PCR(qPCR) and immunocytochemistry.The results showed that：the isolated pSSCs grew well in the feeder layer，and the identification of alkaline phosphatase(AKP) and SOX2，integrinα6，SSEA1，Dazl antibodies were positive，suggesting the pSSCs were in the undifferentiated state.After 8 days of induction，the identification of toluidine blue staining and NSE antibodies were positive，suggesting that the pSSCs had been induced into the neuron-like cells successfully.After 16 days of induction，the identification of ALP，Von Kossa staining and Collagon l antibodies were positive，indicating it had been induced into the osteoblasts.Twenty-two days later，the lipid droplets had been stained into red by oil red O，demonstrated that the pSSCs had been induced into adipocytes.During the process of the pSSCs induction，the results of qPCR demonstrated that，the expression of Nestin and β-tubulin were highest at day 6，the expression of Cbfα1 and Osterix were highest at day 12，however，the mRNA expression of PPARγ and C/EBPα were highest at day 18.In this study，we had obtained the pSSCs successfully，and the pSSCs also had been induced into the neuron-like cells，osteoblasts and adipocytes，separately，comparing the expression of cell-special marker genes.

CLC Number:

S828
S814

CHEN Ting-feng，WANG Xiao-yan，LI Dong，SHI Qing-qing，ZHANG Lei，QIU Feng-long，LIU Zhi-yong，ZHUANG Xun，BIAN Gui-hua，SONG Cheng-yi，LI Bi-chun. Study on the Culture and Induced Differentiation of Spermatogonial Stem Cells in Porcine[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(7): 1104-1112.

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Study on the Culture and Induced Differentiation of Spermatogonial Stem Cells in Porcine

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