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IFN ‐ τ Acts in a Dose‐Dependent Manner on Prostaglandin Production by Buffalo Endometrial Stromal Cells Cultured in vitro
Author(s) -
Chethan SG,
Singh SK,
gsiej J,
Rakesh HB,
Singh RP,
Kumar N,
Agarwal SK
Publication year - 2014
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.12287
Subject(s) - stromal cell , prostaglandin , collagenase , endocrinology , endometrium , medicine , prostaglandin e2 , in vitro , andrology , biology , chemistry , microbiology and biotechnology , enzyme , biochemistry
Contents Interferon‐ τ ( IFN ‐ τ ) has been recognized as the primary embryonic signal responsible for maternal recognition of pregnancy. Uterine endometrium produces both prostaglandin F 2 α ( PGF 2 α ) and prostaglandin E 2 ( PGE 2 ). PGF 2 α is responsible for the luteolysis; however, PGE 2 favours establishment of pregnancy by its luteoprotective action. In this study, the dose‐response effect of recombinant bovine IFN ‐ τ (rb IFN ‐ τ ) on prostaglandin ( PG ) production by buffalo endometrial stromal cells cultured in vitro was studied. Buffalo endometrial stromal cells were isolated by double enzymatic digestion, initially with trypsin III followed by a cocktail of trypsin III , collagenase type II and DN ase I and subsequently cultured till confluence. Further, cells were treated with different doses of rb IFN ‐ τ (0.001, 0.01, 0.1, 1.0 and 10 μg/ml) and keeping a separate set of control. Culture supernatant was collected after 6, 12 and 24 h of treatment. PG levels in the culture supernatant were measured by enzyme immune assay ( EIA ) and total cellular protein estimated by Bradford method. Results indicated that buffalo endometrial stromal cells following rb IFN ‐ τ treatment enhanced the secretion of both PGE 2 and PGF 2 α , and also its ratio in a strict dose‐dependent manner with a significant increase (p < 0.01) in PGE 2 production at 1 μg/ml dose of rb IFN ‐ τ and maximal stimulation for both PG was observed at 10 μg/ml. Further, both PG production and its ratio were increased significantly (p < 0.01) in a time‐dependent fashion in all the groups at 6, 12 and 24 h post‐treatment with highest level achieved at 24 h as compared with control. Absolute levels of PGE 2 remained higher than PGF 2 α indicating PGE 2 as the major PG produced by endometrial stromal cells. The dose‐dependent response of rb IFN ‐ τ signifies the importance of optimum concentration of IFN ‐ τ for the embryonic development especially during the critical period to establish successful pregnancy.