Molecular Nutrition

Isolation and Identification of Duodenal Epithelial Cells of Broiler Embryos in Vitro and Its Establishment and Evaluation of Primary Cultured Absorption Model

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  • Mineral Nutrition Research Division, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received date: 2018-02-06

  Online published: 2018-08-18

Abstract

This experiment was conducted to evaluate the effect of different inoculation density of duodenal epithelial cells on close connectivity and cell viability, and to choose the best cell inoculation density for building absorption model of primary cultured duodenal epithelial cells of broiler embryos in vitro. The experiment was used a single factor complete random design, the duodenal epithelial cells of chick embryos were randomly divided into 3 groups with 6 replicates per group, the inoculation densities were 2.90×106 (group Ⅰ), 6.25×106 (group Ⅱ) and 8.75×106 cells/mL (group Ⅲ), respectively. The experiment lasted for 4 days. The results showed as follows:1) the isolated duodenal epithelial cells were uniform spherical shape, suspended in the medium. After adhered on the wall, the cells began to spread out and attached uniform and had clear boundaries, showing the growth of "paving stones". 2) Duodenal epithelial cells of the positive experimental group were stained blue-black, and the cells of the negative control group were not colored after alkaline phosphatase staining. 3) The transepithelial electrical resistance (TEER) values of cells in groups Ⅰ and Ⅱ were greater than 300 Ω·cm2, and the phenol red penetration rate was less than 5% after 24, 48 or 72 h cultivation. The TEER value of cells in group Ⅰ was significantly higher than that in group Ⅱ (P<0.03), while the phenol red transmittance rate of cells was significantly lower than that in group Ⅱ (P<0.03). 4) The lactate dehydrogenase (LDH) activity of cell culture fluid in group Ⅰ was significantly lower than that in groups Ⅱ and Ⅲ after 24 or 48 h cell cultivation. It is indicated that the cells with inoculation density of 2.90×106 cells/mL have clear boundary, strong growth, close linkage and the best viability after 48 h cultivation. Therefore, the absorption model of primary cultured duodenal epithelial cells in vitro of broiler embryos is established, which provide a model for studying the absorption and molecular mechanism of phosphorus in primary cultured duodenal epithelial cells.

Cite this article

ZHANG Shumin, LIAO Xiudong, LYU Lin, ZAHNG Liyang, LUO Xugang . Isolation and Identification of Duodenal Epithelial Cells of Broiler Embryos in Vitro and Its Establishment and Evaluation of Primary Cultured Absorption Model[J]. Chinese Journal of Animal Nutrition, 2018 , 30(8) : 3159 -3167 . DOI: 10.3969/j.issn.1006-267x.2018.08.034

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