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Culture method of human fallopian tube epithelial cells

An epithelial cell and culture method technology, applied in the field of culturing human fallopian tube epithelial cells, can solve the problem of low cell number, etc., and achieve the effects of mild digestion and high proportion of ciliary differentiation

Inactive Publication Date: 2020-04-28
THE INT PEACE MATERNITY & CHILD HEALTH HOSPITAL OF CHINA WELFARE INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Maobi Zhu et al. proposed a culture method for primary porcine oviduct epithelial cells based on the tracheal epithelial culture model, but the number of cells was small due to the limitation of the culture dish

Method used

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  • Culture method of human fallopian tube epithelial cells
  • Culture method of human fallopian tube epithelial cells
  • Culture method of human fallopian tube epithelial cells

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Embodiment 1

[0043] This embodiment provides a culture method for human fallopian tube epithelial cells, comprising the following steps:

[0044] Step 1, preparation of culture medium and digestion solution

[0045] 1. Basal medium (250ml): Mix DMEM and Ham's F-12 at a volume ratio of 1:1, add 15mM HEPES, 4mM L-glutamine, 3.6mM NaHCO 3 , 100U / ml penicillin and 100μg / ml streptomycin.

[0046] 2. Complete medium (50ml): Add 10 μg / ml insulin, 5 μg / ml transferrin, 0.1 μg / ml cholera toxin, 25ng / ml EGF, 30 μg / ml bovine pituitary extract, 5% Embryonic bovine serum, 0.05 μM retinoic acid, and primary cell antibiotics.

[0047] 3. Serum-free medium (50ml): Add 5 μg / ml insulin, 5 μg / ml transferrin, 0.025 μg / ml cholera toxin, 5 ng / ml EGF, and 30 μg / ml bovine pituitary extract to the basal medium.

[0048] 4. Serum replacement medium (50ml): Add 2% serum replacement to the basal medium.

[0049] 5. Digestive solution: 2700ul Ham F12 (+ L-glutamine + penicillin + streptomycin) + 300ul protease E + 9u...

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Abstract

The invention discloses a culture method of human fallopian tube epithelial cells. The culture method comprises the following steps: obtaining fallopian tube tissues of ampulla, and putting the fallopian tube tissues into digestive juice for digestion; after the digestion is terminated, putting the fallopian tube tissues into a basic culture medium added with 10% fetal bovine serum (FBS) for pre-adherent culture for 3-5 hours; and after the pre-adherent culture, transferring the epithelial cells into a complete culture medium for culture until the cells are dense, then carrying out starvationtreatment for culture for 2 days, and collecting the cells. According to the invention, the fallopian tube tissues are completely digested by specific digestion liquid, and digestion is mild, so thatcell morphology is not damaged; and in addition, the culture medium is optimized in the culture process. According to the culture method, a large number of fallopian tube epithelial cells are obtained, the cilia differentiation ratio is high, and great convenience is provided for related research.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for culturing human fallopian tube epithelial cells. Background technique [0002] The fallopian tubes have long been considered for passive reproduction, serving as sites of fertilization and conduits for the transport of gametes and pre-implantation embryos. Accumulating evidence suggests that the fallopian tube also regulates sperm function and promotes pre-implantation embryo development. The latter functions of the fallopian tubes are mediated by their epithelial cells. The lumen of the oviduct is lined with a columnar epithelium composed of secretory cells and ciliated cells, both of which have critical roles in regulating the epithelial surface of xenogenesis. The mucus produced by secretory cells and the vibration of cilia facilitate the transport of gametes. The proportion of these cell types in different regions of the oviduct changes during the reprodu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2500/25C12N2500/32C12N2500/84C12N2501/01C12N2501/11C12N2501/385C12N2509/00
Inventor 张健张慧宇夏玮赵晓雅黄振齐航朱茜何小青梁桂玲朱晨锋
Owner THE INT PEACE MATERNITY & CHILD HEALTH HOSPITAL OF CHINA WELFARE INST
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