Separation of round spermatids from the rat using an immunoselection panning technique

Molecular Reproduction and Development
S A Pelengaris, H D Moore

Abstract

A method was devised for the isolation of round spermatids from the rat using a positive immunoselection technique (panning). A testis suspension was prepared from adult rats by enzymatic digestion of seminiferous tubules with collagenase. Specific mouse monoclonal antibody (97.25) was indirectly attached to Petri dishes and used in a panning protocol to purify spermatids from the testis cell suspension. The quantity and purity of cells isolated were determined by cell counts and histochemical (periodic acid-Schiff stain) or by immunostaining with acrosome-specific antibodies. A mean yield of 1.38 +/- 0.15 x 10(7) cells per dish was obtained with a purity of more than 90%. The viability of the cells was confirmed by epifluorescent microscopy with propidium iodide/carboxyfluorescein acetate probes. Northern blot analysis of RNA extracted directly from the dish indicated good integrity of a spermatid-specific transcript of glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

References

Jan 1, 1977·Methods in Cell Biology·M L Meistrich
Jan 1, 1990·Journal of Reproduction and Fertility·R A Harrison, S E Vickers
Nov 1, 1991·Molecular Reproduction and Development·Y BlanchardD Lescoat
Feb 1, 1990·Molecular Reproduction and Development·A D'Agostino, M Stefanini
Oct 1, 1989·Biology of Reproduction·K H ThomasM I Simon
Jun 1, 1984·Archives of Biochemistry and Biophysics·J F CrowleyJ Lönngren
Jan 1, 1993·Methods in Enzymology·A R Bellvé

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Citations

Apr 1, 1999·Human Reproduction·B LassalleA Lefèvre
Dec 21, 2013·Zygote : the Biology of Gametes and Early Embryos·Lian ZhuMan-Xi Jiang

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