Effect Of Two-Dimensional And Three-Dimensional Culture On The Gene Expression Of Lactation Metabolism Of Bovine Mammary Epithelial Cells

The main purposes of the experiments was to investigate the effects oftwo-dimensional culture and three-dimensional culture on the gene expression of lactationmetabolism of bovine mammary epithelial cells (BMEC) under different trial conditionwhich included the factors of full nutritional factors, single branched amino acid, hormone.the results were as follows:1. BMEC in Matrigel collagen could rapidly form acinar-like structure, but it formedscattered aggregates in rattail collagen. Furthermore, Matrigel collagen was more stablethan rattail collagen in culture process.The incubation time and the inoculum density ofthree-dimensional culture influenced the gene expression of αs1-casein, β-casein, andκ-casein in BMEC. BMEC of three-dimensional culture formed full ripe acinar-likestructure at5d after incubation, and the optimum inoculum density was1x106/mL.2. On the same nutrients and hormones conditions, the gene expression of αs1-casein,β-casein, and κ-casein in BMEC of three-dimensional culture were significantly higherthan that in two-dimensional culture (P＜0.05), and β-casein in three-dimensional cultureof BMEC could be detected by western blot, but it was not two-dimensional culture ofBMEC.The morphological characters in BMEC of three-dimensional culture were moresimilar to BMEC in vivo.Under two-dimensional culture,the gene expression of αs1-caseinand κ-casein with the addition of prolactin was significantly higher than the control (P＜0.05), the gene expression of α-whey protein in BMEC with the addition of prolactin wassignificantly lower than the control (P＜0.05) and there was no significant difference of thegene expression of β-casein between BMEC with the addition of prolactin and control (P＞0.05) and two-dimensional culture.Under three-dimensional culture,there was nosignificant difference of the gene expression of αs1-casein and κ-casein between BMECwith the addition of prolactin and control(P＞0.05)and the gene expression of β-casein andα-whey protein in BMEC with the addition of prolactin was significantly higher than thecontrol (P＜0.05).3. The gene expression of αs1-casein, β-casein,κ-casein,mTOR, S6K1,4E-BP1, andeIF4E in BMEC by adding leu, ile, and val were significantly higher than the control (P＜0.05)in three-dimensional culture. The conclusion of the experiments was three-dimensional culture of BMEC was muchbetter than that two-dimensional culture of BMEC as in vitro cultivation technique in studyof lactation metabolism of bovine mammary epithelial cells...