Multipotential ability of primitive germ cells from neonatal pig testis cultured in vitro
Multipotential ability of primitive germ cells from neonatal pig testis cultured in vitro
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DOI：
10.1071/rd08176
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发表时间：
2009-01-01
影响因子：
1.9
通讯作者：
Imai, Hiroshi
中科院分区：
文献类型：
--
作者：
Goel, Sandeep;Fujihara, Mayako;Imai, Hiroshi
性细胞是起源于原始生殖细胞的祖细胞类型的生殖细胞，并进一步分化为精原细胞，从而启动精子发生。在本研究中，新分离的性腺细胞有弱表达或无多能决定转录因子的表达，如Pou5f1、SOX2和C-MYC。有趣的是，这些转录因子以及其他重要的转录因子，如NANOG、KLF4和DAZL，在培养细胞中的表达显著上调。原代培养的细胞表达特异的生殖细胞和多能性标志物，如凝集素、扁豆凝集素(DBA)、KIT、ZBTB16、阶段特异性胚胎抗原(SSEA-1)、NANOG和Pou5f1。用单抗特异性鉴定猪生殖细胞，用异种睾丸移植实验测定新鲜细胞和培养细胞的干细胞潜能。仅在移植了新鲜睾丸细胞或原代培养细胞的小鼠睾丸中检测到定植的猪生殖细胞。有趣的是，移植了原代培养细胞的睾丸显示生殖细胞在间质中定植，这反映了它们的致瘤性质。裸鼠皮下注射原代培养的细胞后，从三个生发层产生的组织形成畸胎瘤，证实了它们的多能性。本研究结果可能为建立新生猪睾丸多潜能生殖细胞系提供有用的信息。
Gonocytes are progenitor-type germ cells that arise from primordial germ cells and differentiate further into spermatogonia, thereby initiating spermatogenesis. In the present study, freshly isolated gonocytes were found to have either weak or no expression of pluripotency determining transcription factors, such as POU5F1, SOX2 and C-MYC. Interestingly, the expression of these transcription factors, as well as other vital transcription factors, such as NANOG, KLF4 and DAZL, were markedly upregulated in cultured cells. Cells in primary cultures expressed specific germ cell and pluripotency markers, such as lectin Dolichos biflorus agglutinin (DBA), KIT, ZBTB16, stage-specific embryonic antigen (SSEA-1), NANOG and POU5F1. Using a monoclonal antibody to specifically identify porcine germ cells, the stem cell potential of fresh and cultured cells was determined with a testis xenotransplantation assay. Colonised porcine germ cells were detected only in mouse testes that were either transplanted with fresh testicular cells or with cells from primary cultures. Interestingly, testes transplanted with cells from primary cultures showed colonisation of germ cells in the interstitial space, reflecting their tumourigenic nature. The formation of teratomas with tissues originating from the three germinal layers following the subcutaneous injection of cells into nude mice from primary cultures confirmed their multipotency. The results of the present study may provide useful information for the establishment of multipotent germ stem cell lines from neonatal pig testis.