Isolation of male germ stem cell-like cells from testicular tissue of non-obstructive azoospermic patients and differentiation into haploid male germ cells in vitroopen access
- Authors
- Lee, Dong Ryul; Kim, Kye Seong; Yang, Yun Hee; Oh, Hwa Soon; Lee, Sook Hwan; Chung, Tae Gyu; Cho, Jung Hyun; Kim, Hyun Joo; Yoon, Tae Ki; Cha, Kwang Yul
- Issue Date
- Feb-2006
- Publisher
- OXFORD UNIV PRESS
- Keywords
- embryo production; germ stem cell-like cells; haploid male germ cells; non-obstructive azoospermic patients; spermatogenesis in vitro
- Citation
- HUMAN REPRODUCTION, v.21, no.2, pp.471 - 476
- Indexed
- SCIE
SCOPUS
- Journal Title
- HUMAN REPRODUCTION
- Volume
- 21
- Number
- 2
- Start Page
- 471
- End Page
- 476
- URI
- https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/181798
- DOI
- 10.1093/humrep/dei319
- ISSN
- 0268-1161
- Abstract
- BACKGROUND: The purpose of this study was to establish the culture conditions required to isolate, identify and expand male germ stem cell-like cells (GSC-LC) from the testicular tissue of patients with non-obstructive azoospermia (NOA). METHODS AND RESULTS: Testicular tissues obtained from patients (two with maturation arrest (MA, n = 2) and Sertoli cell-only syndrome (SCOS, n = 11) were dissociated and plated into gelatin-coated dishes. After 2-4 weeks, cultures from both MA patients (100%) and four SCOS patients (36.3%) exhibited multicellular colonies, which proliferated successfully until passage 10. GSC-LC in the colonies displayed alkaline phosphatase activity, as well as Oct-4 and integrin b1 expression after every passage. After the fifth passage, GSC-LC were differentiated by encapsulation in calcium alginate and further cultivation. At 2 and 6 weeks, cells expressed c-Kit, Scp3, testis-specific histone protein 2B (TH2B), and transition protein (TP)-1. Fluorescence in situ hybridization additionally disclosed a few tetraploid and haploid cells at 6 weeks. Human oocytes were activated in the absence of artificial activation and cleaved after the injection of presumptive spermatids. CONCLUSIONS: Our novel culture system may be useful for diagnosing the existence of germ cells and facilitating the treatment of NOA patients.
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