Premium
Practical selection methods for rat and mouse round spermatids without DNA staining by flow cytometric cell sorting
Author(s) -
Hayama Tomonari,
Yamaguchi Tomoyuki,
KatoItoh Megumi,
Ishii Yumiko,
Mizuno Naoaki,
Umino Ayumi,
Sato Hideyuki,
Sanbo Makoto,
Hamanaka Sanae,
Masaki Hideki,
Hirabayashi Masumi,
Nakauchi Hiromitsu
Publication year - 2016
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.22644
Subject(s) - biology , spermatid , flow cytometry , cell sorting , andrology , staining , spermatogenesis , ploidy , reproductive technology , offspring , microbiology and biotechnology , genetics , cryopreservation , gene , embryo , endocrinology , pregnancy , medicine
SUMMARY Round spermatid injection (ROSI) into unfertilized oocytes enables a male with a severe spermatogenesis disorder to have children. One limitation of the application of this technique in the clinic is the identification and isolation of round spermatids from testis tissue. Here we developed an efficient and simple method to isolate rodent haploid round spermatids using flow cytometric cell sorting, based on DNA content (stained with Hoechst 33342 or Dye Cycle Violet) or by cell diameter and granularity (forward and side scatter). ROSI was performed with round spermatids selected by flow cytometry, and we obtained healthy offspring from unstained cells. This non‐invasive method could therefore be an effective option for breeding domestic animals and human male infertility treatment. Mol. Reprod. Dev. 83: 488–496, 2016. © 2016 Wiley Periodicals, Inc .