Journal
STAR PROTOCOLS
Volume 3, Issue 3, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.xpro.2022.101540
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Funding
- Ministry of Science and Technology of China [2019YFA0802600]
- Postdoctoral Research Foundation of China [2020M671885]
- National Natural Science Foundation of China [31970793, 32170856]
- Fundamental Research Funds for the Central Universities [WK2070000156]
- USTC [KY9100000001]
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The study introduces an optimized centrifugal elutriation protocol for rapid and accurate isolation of mouse germ cells at different stages, providing an important tool for studying germline development.
Germline development is challenging to study due to the diversity of cell types in mammalian testis. Here, we present an optimized protocol, namely centrifugal elutriation, that allows the simultaneous isolation of mouse germ cells at different stages with high purity within & DBLBOND;2 h. This approach exploits the JE-5.0 centrifugal elutriation system that fractionates cells based on differential sedimentation gravity. We herein provide the optimized parameters and procedures for isolation of elongating spermatids, round spermatids, and pachytene spermatocytes from adult mouse testes. For complete details on the use and execution of this protocol, please refer to Bao et al. (2018).
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