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Method for culturing endometrial cells

A technology of endometrium and cells, applied in the biological field, can solve the problems of slow growth and poor passage effect

Pending Publication Date: 2021-08-10
博品(上海)生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

As we all know, the culture method has a decisive relationship with the ability of the cells to be subcultured and the viability. The existing technology generally uses basal medium, fetal bovine serum, penicillin, and streptomycin as the culture system, and the cell density reaches 80%. And many problems such as poor subculture effect (Mol Hum Reprod, 2013, 19 (6): 361-368)

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  • Method for culturing endometrial cells
  • Method for culturing endometrial cells
  • Method for culturing endometrial cells

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1 A method for culturing endometrial cells, the steps are as follows:

[0026] Add 5 mg of stem cell exosome freeze-dried powder to 5 mL of ultrapure water to dissolve, then add 100 mL of DMEM / F12 medium, and then add 5 mL of human albumin (the volume of human serum albumin is the volume of DMEM / F12 medium 5%) to obtain a cell culture system.

[0027] Take 1 ml of cell suspension, add 10 mL of cell culture system, mix, and centrifuge at 1500 rpm for 5 minutes, discard the supernatant, and resuspend the obtained cell pellet to 1 ml with cell culture system, follow 10 4 / cm 2 Inoculate culture flasks at a density of 100%, add them to T150 cell culture flasks, and add 25ml of cell culture system to each culture flask, as the 0th day of cell growth, at 37°C, 5% CO 2 Culture in an incubator, update the cell culture system every 3 days, after the cells grow to the 7th day, the cell density reaches 90%, the cells can be subcultured, the cells are photographed, and...

Embodiment 2

[0036] Add 1 mg of stem cell exosome freeze-dried powder to 5 mL of ultrapure water to dissolve, then add 100 mL of DMEM / F12 medium, and then add 5 mL of human albumin (the volume of human serum albumin is the volume of DMEM / F12 medium 5%) to obtain a cell culture system. Take 1 ml of cell suspension, add 10 mL of cell culture system, mix, and centrifuge at 1500 rpm for 5 minutes, discard the supernatant, and resuspend the obtained cell pellet to 1 ml with cell culture system, follow 10 4 / cm 2 Inoculate culture flasks at a density of 100%, add them to T150 cell culture flasks, and add 25ml of cell culture system to each culture flask, as the 0th day of cell growth, at 37°C, 5% CO 2 The cell culture system was updated every 3 days. After the cells grew to the 7th day and the cell density reached 75%, the cells were photographed to observe the cell growth status. In addition, after 7 days of culture, the absorbance of 12 hours was 1.062.

Embodiment 3

[0038] Add 10 mg of stem cell exosome freeze-dried powder to 5 mL of ultrapure water to dissolve, then add 100 mL of DMEM / F12 medium, and then add 5 mL of human albumin (the volume of human serum albumin is the volume of DMEM / F12 medium 5%) to obtain a cell culture system. Take 1 ml of cell suspension, add 10 mL of cell culture system, mix, and centrifuge at 1500 rpm for 5 minutes, discard the supernatant, and resuspend the obtained cell pellet to 1 ml with cell culture system, follow 10 4 / cm 2 Inoculate culture flasks at a density of 100%, add them to T150 cell culture flasks, and add 25ml of cell culture system to each culture flask, as the 0th day of cell growth, at 37°C, 5% CO 2 Cultured in an incubator, and the cell culture system was updated every 3 days. After the cells grew to the 7th day and the cell density reached 80%, the cells were photographed to observe the growth status of the cells. In addition, after 7 days of culture, the absorbance at 12 hours was 1.657. ...

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Abstract

The invention discloses a method for culturing endometrial cells, which comprises the following steps: adding a stem cell exosome freeze-dried powder aqueous solution and human serum albumin into a DMEM / F12 culture medium to obtain a cell culture system; resuspending endometrial cells obtained by separating endometrial tissues by using a DMEM / F12 culture medium, adding the resuspended endometrial cells into the cell culture system, then carrying out centrifugal separation, taking precipitates, resuspending the precipitates by using the cell culture system, and inoculating a culture bottle for culturing to complete the culture of the endometrial cells. According to the method, when the cells grow to the seventh day, the cell density reaches 90%, cell passage can be carried out, and the cell activity is good.

Description

technical field [0001] The invention belongs to biotechnology, and in particular relates to a method for culturing endometrial cells. Background technique [0002] The endometrium is composed of a single layer of columnar epithelium and the lamina propria. The etiology of endometrial injury includes postpartum curettage, spontaneous abortion, induced abortion, endometrial ablation, pelvic tuberculosis, and infection. Destruction of the myometrium, research suggests that when the basal layer of the endometrium is damaged, it may lead to the loss or damage of endometrial cells. The existing technology can effectively separate cells from tissues, then culture them, and then reinfuse them into autologous bodies to achieve damage repair. In addition, the cultured cells can be used as objects to detect antigens (EMAg), reducing the need for human materials. The cell culture process is complicated, and the prior art uses the basal medium as a necessary component, adding appropriat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2500/84C12N2501/998C12N2509/00
Inventor 董健伸徐汝强孙永沛程洪斌王晓东孟凡江
Owner 博品(上海)生物医药科技有限公司
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