Prolonged Survival of Cock Spermatozoa In Vitro with Fluid Removed from Tissue Cultured Oviducal Cells

Abstract

When cock spermatozoa were incubated at 41 C with fluid removed from tissue cultured oviducal cells, their motility, assessed at room temperature, was maintained for 4 or 5 days. Fertilizing ability was retained for one week after culture for two days at 41 C. These results were comparable to the duration of motility and fertilizing ability of spermatozoa incubated in the presence of cultured oviducal cells. The results indicated that the fluid removed from the tissue cultured cells contains a secretory product(s) capable of prolonging in vitro the motility and fertilizing ability of spermatozoa.

Preservation of the Fertilizing Capacity of Cock Semen Incubated In Vitro at 41 C

“…Considering the fertility results for the three groups of spermatozoa incubated for either 6 or 24 hr, MEM alone appears to be as beneficial as TCF or the presence of TCC in maintaining in vitro sperm survival at 41 C. This observation is contrary to those of Ashizawa et al (1976), Ashizawa and Nishiyama (1977a), and Fujihara and Howarth (1980), all of whom reported that spermatozoa incubated in MEM alone had little or no fertilizing ability following surgical insemination. An explanation for these contrasting results may be the longer sperm incubation periods of 2 and 4 days used in these studies as compared to the shorter incubation periods of 6 and 24 hr used in the present studies.…”

“…Cells to be cultured were obtained from whole 9-day-old chick embryos. Cell preparation and culture were carried out according to the method described by Fujihara and Howarth (1980). The culture medium referred to as minimum essential medium (MEM) consisted of nine parts MEM (Eagle modified autoclavable medium with Earle's salts-#F-17 obtained from Grand Island Biological Company) and one part heat inactivated (56 C for 30 min) fetal calf serum.…”

“…Subsequent studies by Ashizawa and Nishiyama (1977a,b;1978) indicated that the effectiveness of this system depended upon the close association of the spermatozoa with the tissue cultured cells during incubation. Fujihara and Howarth (1980) demonstrated that tissue culture fluid removed from cultured cells (obtained from the shell gland) was capable of maintaining sperm survival. The major limitations were: 1) the small number of cells that could be harvested from the shell gland for establishing monolayers and 2) the small numbers of spermatozoa that could be incubated at any one time and 3) the requirement of surgical insemination to evaluate fertility.…”

Oviducal storage of spermatozoa in the turkey: Its relevance to artificial insemination technology

“…They have since reported that fowl spermatozoa can survive at 41°C for 5 to 6 d when co-cultured either with dispersed somatic cells from various tissues from different species (Ashizawa et al, 1982 or for up to 11 d in expiants of oviducal tissue . It has also been reported that fowl spermatozoa can survive up to 4 d in cell-free media recovered from cell cultures (Fujihara and Howarth, 1980;Fujihara and Koga, 1982). Because cells and their products appear to maintain spermatozoal viability, the use of cell culture for short-term storage of semen is intriguing.…”

Scanning Electron Microscopy of Fowl Oviducal Cells and HeLa and BHK-21 Cells Incubated with or Without Fowl Spermatozoa In Vitro

“…Since then, fowl spermatozoa incubated with various types of cultured cells, such as cells from the fowl kidney, the skeletal muscle of a 9-day chick embryo, and HeLa and BHK-21 cells, survived for approximately 5 to 7 days, even at 41 C (Ashizawa and Nishiyama, 1977;Fujihara and Howarth, 1980;Ashizawa et al, 1982).…”