In vitro-cultured bovine granulosa and oviductal cells secrete sperm motility-maintaining factor(s).
Since bovine cumulus oophorous and oviductal 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 oviductal 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.(ABSTRACT TRUNCATED AT 250 WORDS)
Ijaz
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A
Departement de Zootechnie, FSAA, Université Laval, Québec, Canada.
Lambert
R D
RD
Sirard
M A
MA
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D005187:Fallopian Tubes / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D005260:Female; D015571:Follicular Fluid / Q000166:cytology / Q000502:physiology; D006107:Granulosa Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D008297:Male; D013081:Sperm Motility
D017077:Culture Media, Conditioned