Aarti Sukanta Mondal
ICAR-NIANP, India
Title: Modulation of growth pattern in buffalo endometrial epithelial and stromal cells by LPS and TNFα
Biography
Biography: Aarti Sukanta Mondal
Abstract
Statement of the Problem: Early embryonic loss is one of the major factors affecting reproductive efficiency in buffalo. Prostaglandins are mediators of various reproductive events viz., luteolysis, implantation and parturition in farm animals. LPS and TNFα play a crucial role in regulation of prostaglandin production by endometrial cells by modulating the growth pattern of epithelial and stromal cells. The objective of the study was to test the effect of LPS and TNFα on in vitro growth pattern of endometrial epithelial and stromal cells.
Methodology & Theoretical Orientation: Endometrial epithelial cells were isolated by infusing HBSS containing 0.3% trypsin into mid luteal uterine horn and were cultured in RPMI 1640 medium at 38.5 oC in presence of 5% CO2 for 7 days. For isolation of stromal cells, endometrial tissues were digested at 37 oC for 1 hour in RPMI-1640 medium containing collagenase. The contaminating epithelial cells were eliminated by treatment with 0.25% trypsin solution after 16 hours of culture and the remaining cells were cultured in RPMI 1640 medium at 38.5 oC in presence of 5% CO2 for 7 days. After reaching the confluency, the endometrial epithelial and stromal cells were treated with increasing doses of LPS (1, 10,100, 1000, 104, 105 ng) and TNFa (0.005, 0.05, 0.5, 1, 2.5, 5 nM) for 24 hour.
Findings: In primary culture, epithelial cells exhibited cuboidal or columnar morphologies and showed contact inhibition at the stage of confluence. In primary culture, stromal cells exhibited flat, fibroblastic and spindle shaped and showed streaming at the stage of confluence. The protein content of endometrial epithelial cells was lowest in control group which increased a maximum level at 105 ng LPS in endometrial epithelial cells. In stromal cells, protein content increased to reach a maximum level at 100 ng LPS which then declined to lowest level at 105 ng LPS. Protein concentration was found maximum in the control group of endometrial epithelial cells but lowest level was observed at 2.5 nM TNF-a. In stromal cells, protein content was lowest at 0.005 nM TNF-a which then increased to a nadir at 1 nM dose of TNF-a. The concentration of protein was highest in control group and lowest level was at 2.5 nM TNF-a.
Conclusion & Significance: It can be concluded that the growth pattern of endometrial epithelial and stromal cells was modulated by LPS and TNF-a in vitro.