Long-term culture and transplantation of murine testicular germ cells
| Autor: | Dongkee Jeong, Michael D. Griswold, Derek J. McLean |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Male
medicine.medical_specialty Urology Endocrinology Diabetes and Metabolism Basic fibroblast growth factor Cell Culture Techniques Stem cell factor Mice Inbred Strains Mice Transgenic Biology Andrology chemistry.chemical_compound Mice Endocrinology Germ cell proliferation Internal medicine Testis medicine Animals Oligospermia Spermatozoa Transplantation medicine.anatomical_structure Reproductive Medicine chemistry Subculture (biology) Leukemia inhibitory factor Totipotent Stem Cells Germ cell Fetal bovine serum Cell Division Stem Cell Transplantation |
| Zdroj: | Journal of andrology. 24(5) |
| ISSN: | 0196-3635 |
| Popis: | The objectives of this study were to develop an in vitro culture system to optimize germ cell proliferation and to measure the potential of the cultured germ cells to produce mature spermatozoa after transplantation into a recipient. Donor germ cells isolated from ROSA26 male mice were cultured with a STO feeder cell layer in Dulbecco's minimal essential medium (DMEM) supplemented with fetal bovine serum (FBS), stem cell factor, leukemia inhibitory factor, basic fibroblast growth factor, insulin-like growth factor 1, interleukin-11, L-glutamine, sodium pyruvate, 2-mercaptoethanol, murine oncostatin M, and platelet-derived growth factor. Donor germ cells formed colonies in the primary cultures after 8-21 days. These cultured colonies were maintained for 4 weeks or longer without subculture and proliferated for up to 8 passages over a period of 3 months. These colonies had alkaline phosphatase activity and incorporated 5-bromo-2'-deoxyuridine. These colonies were positive partially when screened with antibody for germ cell nuclear antigen and c-kit. Germ cells cultured with this supplemented medium showed enhanced colonization vs controls cultured with DMEM and FBS. Cultured germ cells from Rosa26 donors were transplanted into testes and were identified by X-gal staining and histological screening. The cells cultured in the supplemented medium colonized the tubules and initiated spermatogenesis in the recipient mice. This is an improved method for culturing germ cells and may be useful in gene therapy and the production of transgenic animals. |
| Databáze: | OpenAIRE |
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