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期刊名称: Biology of Reproduction
Volume:82    Issue:2        Page:363-372
ISSN:0006-3363

Isolation, Characterization, and Culture of Human Spermatogonia期刊论文

作者: Zuping He Maria Kokkinaki Jiji Jiang Ina Dobrinski Martin Dym
DOI:10.1095/biolreprod.109.078550

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页码: 363-372
被引频次: 161
出版者: Society for the Study of Reproduction,SOC STUDY REPRODUCTION
期刊名称: Biology of Reproduction
ISSN: 0006-3363
卷期: Volume:82    Issue:2
语言: English
摘要: This study was designed to isolate, characterize, and culture human spermatogonia. Using immunohistochemistry on tubule sections, we localized GPR125 to the plasma membrane of a subset of the spermatogonia. Immunohistochemistry also showed that MAGEA4 was expressed in all spermatogonia (A dark , A pale , and type B) and possibly preleptotene spermatocytes. Notably, KIT was expressed in late spermatocytes and round spermatids, but apparently not in human spermatogonia. UCHL1 was found in the cytoplasm of spermatogonia, whereas POU5F1 was not detected in any of the human germ cells. GFRA1 and ITGA6 were localized to the plasma membrane of the spermatogonia. Next, we isolated GPR125-positive spermatogonia from adult human testes using a two-step enzymatic digestion followed by magnetic-activated cell sorting. The isolated GPR125-positive cells coexpressed GPR125, ITGA6, THY1, and GFRA1, and they could be cultured for short periods of time and exhibited a marked increase in cell numbers as shown by a proliferation assay. Immunocytochemistry of putative stem cell genes after 2 wk in culture revealed that the cells were maintained in an undifferentiated state. MAPK1/3 phosphorylation was increased after 2 wk of culture of the GPR125-positive spermatogonia compared to the freshly isolated cells. Taken together, these results indicate that human spermatogonia share some but not all phenotypes with spermatogonial stem cells (SSCs) and progenitors from other species. GPR125-positive spermatogonia are phenotypically putative human SSCs and retain an undifferentiated status in vitro. This study provides novel insights into the molecular characteristics, isolation, and culture of human SSCs and/or progenitors and suggests that the MAPK1/3 pathway is involved in their proliferation.
相关主题: A SPERMATOGONIA, G protein-coupled receptor 125 (GPR125), MOUSE, C-KIT, isolation, testis, LINE STEM-CELLS, human putative SSCs and progenitor cells, SEMINIFEROUS EPITHELIUM, REPRODUCTIVE BIOLOGY, CELL SELF-RENEWAL, culture, spermatogenesis, phenotypic characterization, MAPK1/3 pathway, human spermatogonial stem cells, DIFFERENTIATION, SIGNAL-REGULATED KINASE, RHESUS MACAQUES, Immunohistochemistry, Spermatocytes - chemistry, Spermatogonia - classification, Humans, Middle Aged, Male, Stem Cells - cytology, Cell Membrane - chemistry, Cell Division, Adult, Antigens, Neoplasm - analysis, Cell Culture Techniques, Spermatogonia - cytology, Receptors, G-Protein-Coupled - analysis, Spermatogonia - chemistry, Testis - cytology, Cell Separation, Biomarkers - analysis, Stem Cells - chemistry, Phenotype, Mitogen-Activated Protein Kinase 3 - metabolism, Adolescent, Neoplasm Proteins - analysis, Spermatids - chemistry, Enzyme Activation, Microscopy, Fluorescence, Mitogen-Activated Protein Kinase 1 - metabolism, Index Medicus,

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