Evaluation of Culture Time and Media in anIn VitroTestis Organ Culture System
| Autor: | Takehiko Ogawa, Daniel T. Sloper, Noriko Nakamura, Deborah K. Hansen, Takuya Sato, Gwenn E. Merry, Pedro L. Del Valle, Amy L. Inselman |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Embryology 030219 obstetrics & reproductive medicine Spermatid Health Toxicology and Mutagenesis Spermatid differentiation Biology Toxicology Organ culture Sperm In vitro Andrology 03 medical and health sciences 030104 developmental biology 0302 clinical medicine medicine.anatomical_structure In vivo Pediatrics Perinatology and Child Health Immunology medicine Spermatogenesis Developmental Biology Explant culture |
| Zdroj: | Birth Defects Research. 109:465-474 |
| ISSN: | 2472-1727 |
| Popis: | Background The complexity of spermatogenesis makes development of appropriate in vitro testis models challenging. A novel in vitro mouse testis culture system has been reported but not yet evaluated as an alternative model for male reproductive toxicity testing. We assessed the effects of media composition on sperm differentiation and testis morphology of cultured mouse testis fragments. Methods Testes from postnatal day 5 B6:CBA-Tg(Acrv1-EGFP)2727Redd/J male mice were cultured in knockout serum replacement (KSR) or Albumax I (Albumax) medium. Enhanced green fluorescent protein (EGFP) expression was examined on days 35, 42, 45, and 49 of culture. Histology and flow cytometry were performed for testis morphology and spermatid differentiation. Results EGFP signals were first observed in round spermatids on day 22 of culture (corresponding to postnatal day 27) and were observed until the end of culture, indicating testis-specific protein expression. A-kinase anchor protein 4 expression, a marker of elongated spermatid (step 15–16) occurred earlier in explants cultured in KSR than Albumax medium (typically day 35 and after day 42 of culture, respectively). The percentage of seminiferous tubules with elongated spermatid was higher in Albumax than KSR medium from days 45 to 49 of culture. Conclusion Albumax medium may facilitate or support better morphology and spermatid production than KSR medium. Further studies need to improve spermatid production and refinement of this in vitro testis culture system that may be useful as a supplement to current male reproductive toxicity testing or an alternative model in cases where in vivo testing may be unfeasible. Birth Defects Research 109:465–474, 2017. © 2017 Wiley Periodicals, Inc. |
| Databáze: | OpenAIRE |
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