Purification and embryotropic roles of tissue inhibitor of metalloproteinase-1 in development of "HanWoo" (Bos taurus coreanae) oocytes co-cultured with bovine oviduct epithelial cells.
This study was conducted to purify a tissue inhibitor of metalloproteinase (TIMP)-1 in a serum-free medium conditioned with bovine oviduct epithelial cells (BOEC) and to evaluate its effect on development of "HanWoo" (Bos taurus coreanae) embryos to the blastocyst stage. In the first study using SDS-PAGE electrophoresis, the presence of 32 kDa proteins, which contains TIMP-1, was detected in the medium conditioned with BOEC, and TIMP-1 was then purified from the medium by gel filtration and HPLC techniques. When examined TIMP-1 secretion, fluorescent foci indicating the secretion of TIMP-1 were found after stained BOEC with fluorescein isothiocyanate. In the next experiment, two-cell embryos derived from in vitro-fertilization were cultured in a serum-free medium, to which 0, 1.25, 2.5 or 5 microg/ml of purified TIMP-1 was supplemented. More (P<0.05) embryos developed to the morula and blastocyst stages after the addition of 2.5 microg/ml to culture medium than after no addition. In conclusion, our data indicate that BOEC secrete TIMP-1 and this glycoprotein promotes the prehatched development of "HanWoo" embryos derived from in vitro-fertilization.
Hwang
W
W
Department of Theriogenology, College of Veterinary Medicine, Seoul National University, Korea.
Kim
H
H
Lee
E
E
Lim
J
J
Roh
S
S
Shin
T
T
Hwang
K
K
Lee
B
B
D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle / Q000196:embryology* / Q000502:physiology; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000662:veterinary; D002852:Chromatography, Ion Exchange / Q000662:veterinary; D018920:Coculture Techniques / Q000662:veterinary; D017077:Culture Media, Conditioned; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology; D005187:Fallopian Tubes / Q000166:cytology / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D008297:Male; D009865:Oocytes / Q000502:physiology*; D011480:Protease Inhibitors / Q000302:isolation & purification*; D011817:Rabbits; D011897:Random Allocation; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology
D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; D011480:Protease Inhibitors; D019715:Tissue Inhibitor of Metalloproteinase-1