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In vitro‐cultured bovine granulosa and oviductal cells secrete sperm motility‐maintaining factor(s)
Author(s) -
Ijaz Ahmad,
Lambert Raymond D.,
Sirard MarcAndre
Publication year - 1994
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.1080370108
Subject(s) - motility , biology , andrology , bovine serum albumin , sperm motility , sperm , follicular fluid , fetal bovine serum , centrifugation , secretion , in vitro , endocrinology , medicine , immunology , embryo , biochemistry , oocyte , microbiology and biotechnology
Since bovine cumulus oophorous and oviductual cell cultures are known to support and maintain frozen‐thawed bovine sperm viability and motility for extended time periods, we investigated whether granulosa cell (GC)‐ and oviductual cell (OC)‐conditioned media have similar effects. GC and OC were cultured for 3 days in TCM‐199 medium supplemented with 10% fetal calf serum. At that time, the supernatant was discarded from GC and the monolayers were covered with Sp‐TALP medium containing 6 mg/ml bovine serum albumin, while the OC were recovered by centrifugation and transferred to culture bottles containing Sp‐TALP. Two days later, GC‐conditioned and OC‐conditioned Sp‐TALP were recovered and dialyzed, and their retentates were lyophilized. Bovine follicular fluid (BFF) was also dialyzed, and its retentate was lyophilized. When sperm were incubated in GC‐ or OC‐conditioned media, motility remained above 62% and 42% at 6 hr and 30 hr, respectively, and motility was higher than that of the control both at 6 hr (39%; P < 0.001) and at 30 hr (9%; P < 0.0001). Similarly, when sperm were incubated in the lyophilized retentates of GC‐ and OC‐conditioned media and in BFF at a dose of 0.1, 0.5, or 1.0 mg/ml, the motility rates were higher both at 6 hr ( P < 0.05) and at 30 hr ( P < 0.01) compared to the control. The increase in motility was dose dependent; a 1.0 mg/ml dose improved ( P < 0.05) motility compared to a 0.1 mg mg/ml dose. Heat treatment of the retentates of GC, OC, and BFF at 55°C for 30 min did not destroy their ability to support and maintain motility. However, heating at 100°C for 5 min destroyed their ability to support motility. Molecular sieving of retentates on Sephancryl S‐300 yielded fractions that were highly effective ( P < 0.01) in enhancing and maintaining motility compared to the other fractions. In conclusion, GC and OC secrete nondialyzable, heat‐labile factor(s), which support and maintain sperm viability and motility for up to 30 hr. © 1994 Wiley‐Liss, Inc.