Preparation of spermatogonia, spermatocytes, and round spermatids for analysis of gene expression using fluorescence-activated cell sorting.
A new method is described for the purification of spermatogenic cell populations from mouse testis. Through use of this method, it is possible to purify leptotene, zygotene, and pachytene primary spermatocytes as well as round spermatids from adult mouse testis. In addition, spermatogonial populations can be purified from mice at 9 days postpartum. The leptotene and zygotene primary spermatocytes that can be prepared by this method are impossible to separate successfully by the unit gravity method. The cells were used to prepare RNA for reverse transcriptase-polymerase chain reactions.
Division of Biology, Beckman Research Institute, City of Hope National Medical Center, Duarte, California 91010, USA.
L L
LL
Mays-Hoopes
J
J
Bolen
A D
AD
Riggs
J
J
Singer-Sam
D000818:Animals; D002469:Cell Separation; D005434:Flow Cytometry; D015870:Gene Expression; D008297:Male; D008540:Meiosis; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification; D012194:RNA-Directed DNA Polymerase; D013087:Spermatids / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D013090:Spermatocytes / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D013093:Spermatogonia / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D013737:Testis / Q000166:cytology
D012313:RNA; D012194:RNA-Directed DNA Polymerase