A kind of in vitro culture method of mouse fallopian tube epithelial cells

An epithelial cell, in vitro culture technology, applied in artificial cell constructs, animal cells, vertebrate cells, etc., can solve the problem of many passages of fallopian tube epithelial cells, decreased cell activity and secretory function, biological activities such as loss of cilia swing, etc. question

Active Publication Date: 2017-08-29
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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  • Claims
  • Application Information

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Problems solved by technology

However, this treatment will inevitably lead to excessive passage times of fallopian tube epithelial cells, reduce cell viability and secretion function, and lose some characteristic biological activities such as cilia swinging.

Method used

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  • A kind of in vitro culture method of mouse fallopian tube epithelial cells

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Embodiment 1

[0015] (1) Preparation of culture medium

[0016] Mix 50 milliliters of FBS, 2.5 milliliters of penicillin-streptomycin mixed solution [penicillin-streptomycin mixed solution (100X), the content of penicillin is 10000 U / ml, the content of streptomycin is 10 mg / ml], 0.15 gram of sodium pyruvate, 100 Microliter of epidermal growth factor (EGF, dissolve 200 micrograms of EGF in 2 ml of five-distilled water, store at -20°C for later use), 50 microliters of type I collagen solution (C8919, Sigma, USA; dissolve 50 mg of collagen powder in 8 ml of five-distilled In water, add acetic acid dropwise to adjust the pH to 3.0, and after fully dissolving, adjust the volume to 10 ml with five-distilled water) Dissolve in 450 ml of DMEM / F12 culture solution, filter through a 0.22 micron filter membrane, and dispense into sterile 50 40 ml per tube and store at 4°C. Preheat one tube of culture medium in a 37°C water bath half an hour before cell culture.

[0017] (2) Syringe handling

[0018...

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Abstract

The invention discloses an in-vitro culture method of mouse oviduct epithelial cells. A mouse oviduct is placed in a plate along with a uterine horn end, pancreatic enzyme digestive juice is injected from into the uterine horn end through a syringe with a blunt needle to be digested for 5 to 8 minutes at 37 DEG C, the uterine horn end is cut off, the oviduct is gently squeezed through ophthalmic forceps, mucosal tissue and cells which flow out are collected in a centrifuge tube to be centrifuged, supernatant is removed and a culture solution is added, the mucosal tissue and cells are inoculated into a culture bottle after blowing, beating and mixing to be cultured in an energy substance sodium pyruvate, epidermal growth factor and collagen contained culture solution, cells start to grow adhering to the wall after 18 to 24 hours, and about 80 to 90% of cells adhere to the wall after 4 to 5 days, wherein the significant promotion effect on the epithelial cell proliferation is achieved through the energy substance sodium pyruvate, cultivation microenvironment is adjusted through epidermal growth factors, and the cell adherence is facilitated through collagen.

Description

technical field [0001] The invention relates to a biological culture technology, in particular to a method for in vitro culture of mouse oviduct epithelial cells. Background technique [0002] For mammals, the fallopian tube is the place where many important reproductive events occur, including egg maturation, sperm selection and storage, sperm-egg union, early embryo development, and the first close contact between the embryo and the mother, etc. It plays an important role in the study of its regulation. Studies have shown that the fallopian tube epithelial cells, as the first barrier for material exchange and communication with gametes or early embryos, can effectively simulate the fallopian tube environment in vivo, and are very suitable for studying reproductive events in early pregnancy. At present, the in vitro culture system of oviduct epithelial cells of larger mammals such as cattle, goats, humans, and pigs has been successfully established. However, as an importan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
Inventor 谭秀文靳青游伟成海建刘倚帆刘晓牧刘桂芬万发春宋恩亮
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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