RRC ID 76664
Author Hashimoto K, Odaka H, Ishikawa-Yamauchi Y, Nagata S, Nakamura H, Kimura H, Sato T, Makiyama K, Ogawa T.
Title Culture-space control is effective in promoting haploid cell formation and spermiogenesis in vitro in neonatal mice.
Journal Sci Rep
Abstract The classical organ culture method, in which tissue is placed at the gas‒liquid interphase, is effective at inducing mouse spermatogenesis. However, due to reginal variations in the supply of oxygen and nutrients within a tissue, the progress of spermatogenesis was observed only in limited areas of a tissue. In addition, haploid cell formation and its differentiation to spermatozoon, i.e. spermiogenesis, were infrequent and inefficient. Here, we show that the polydimethylsiloxane (PDMS)-chip ceiling (PC) method, which ensures a uniform supply of nutrients and oxygen throughout the tissue by pressing it into a thin, flat shape, can provide control over the culture space. We used this method to culture testis tissue from neonatal mice, aged 1 to 4 days, and found that modulating the culture space during the experiment by replacing one chip with another that had a higher ceiling effectively increased tissue growth. This adjustment also induced more efficient spermatogenesis, with the process of spermiogenesis being particularly promoted. Meiotic cells were observed from culture day 14 onward, and haploid cells were confirmed at the end of each experiment. This technique was also shown to be a sensitive assay for testicular toxicity. Culture-space control will be a critical regulation parameter for sophisticated tissue culture experiments.
Volume 13(1)
Pages 12354
Published 2023-7-31
DOI 10.1038/s41598-023-39323-y
PII 10.1038/s41598-023-39323-y
PMID 37524742
PMC PMC10390558
MeSH Animals Animals, Newborn Haploidy Male Mice Spermatogenesis* / physiology Spermatozoa Testis*
IF 3.998
Mice RBRC00886