Effects Of Bovine Peripheral-Blood Lymphocyte On The Apoptosis And Proliferation Of Oviduct Epithelium Cells And Granulose Cells

The objective of this study is to investigate the effects of bovine PBL on theapoptosis and proliferation of oviduct epithelium cells(OEC) and granulosecells(GCs). Isolated peripheral-blood lymphocyte(PBL) was co-cultured with the sameamount of purified oviduct epithelial cells, granulose cells at the ratio of 10:1, 5:1, 3:1, 1:1for 4 days, respectively, in 24-well culture dish, and the oviduct epithelial cells andgranulose cells treated as control. The basic growth condition of oviduct epithelial cellswas observed a under converted microscope, so did the survival rate of cells stained withtrypan blue and cells treated with PI. The cell cycle stage and apoptosis of oviductepithelial cells and mural granulose cells were detected by flow cytometry. The resultswere as follow. When OEC and PBL were co-cultured, PBL promote OECapoptosis. There is differ significantly between every test group and control group. The ratio ofapoptosis of 5:1 group is highest and 10:1 group is lowest. There is no differ significantlybetween 3:1 group and 1:1 group(P＞0.05). Each of remained groups is differsignificantly. The results indicated that the ratio of OEC and PBL decided OEC apoptosis;When GCs and PBL are co-cultured, PBL inhibit GCs apoptosis. The ratio of granulosecells apoptosis of 5:1 group and 1:1 group is lower than control group. There is differsignificantly between 10:1 group and 3:1 group, then high differ significantly between 10:1 group andcontrol group. There is no differ significantly in the middle of 5:1group, 1:1group andcontrol group(P＞0.05).To investigate the effect of PHA added into the co-culture system on the apoptosis ofoviduct epithelial cells and granulose cells, preliminary experiments were conducted toexplore the effect of PHA on the apoptosis of oviduct epithelial cells and mural granulosecells. The results indicated that PHA had no obvious effect on the growth and apoptosisof GCs(P＞0.05), but improved the proliferation of oviduct epithelial cells(P＜0.05).Based on the co-culture results, the peripheral-blood lymphocytes were co-cultured withoviduct epithelial cells at passage 4 and 8 at the ratio of 5:1 for 4 days in mediumsupplemented with PHA, and were detected by flow cytometry for the apoptosis of oviduct epithelial cells after stained by Hochest 33342/PI. The results were as follows:the apoptosis of oviduct epithelial cells at different passages was quite low, which wasclose to that of control group. In other words, It is presumed PHA added in co-culturemedium inhibited the apoptosis of oviduct epithelial cells.