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Perfusion device and perfusion method for embryonic stem cell culture

An embryonic stem cell and stem cell technology, applied in the field of stem cells, can solve the problems of stem cell consumption exceeding the extraction amount, medium loss, and short supply.

Inactive Publication Date: 2021-07-23
姜胜维
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Stem cell research provides protection for people's health. Stem cells, as ancestor cells, can differentiate into various cells in the human body, which has a strong therapeutic guarantee for cancer or various serious diseases, but stem cells need to be extracted. When cultivating and treating cancer or serious diseases, the amount of stem cells required is relatively large, so if stem cells are not cultivated, the consumption of stem cells is much higher than the amount extracted, resulting in a situation in which the supply exceeds demand
[0003] At present, in the process of stem cell culture, it is generally carried out in the form of perfusion. Perfusion is to place the stem cells in the culture container, and continuously supply the medium through the connecting tube. The used medium is taken out of the culture container to continuously provide medium for stem cells. However, in the process of culturing stem cells, stem cells require different amounts of medium at different maturation stages. If the amount of supply medium remains unchanged, it will Cause unnecessary loss of medium, so we introduced a perfusion device and perfusion method for stem cell culture to solve such problems

Method used

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  • Perfusion device and perfusion method for embryonic stem cell culture
  • Perfusion device and perfusion method for embryonic stem cell culture
  • Perfusion device and perfusion method for embryonic stem cell culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A perfusion method for stem cell culture, specifically as follows:

[0037] S1, preparation work, prepare the stem cell culture medium, and lower the temperature of the stem cell culture medium to 6°C;

[0038] S2, put the stem cells into the container, and place it in the perfusion box 5;

[0039] S3, in the perfusion stage, connect the stem cell culture medium to the feed pipe 3, and enter the perfusion box 5 through the discharge pipe 4;

[0040] S4, turn on the peristaltic pump in the perfusion box 5, and keep the temperature at 30°C;

[0041] S5, ventilate oxygen, give perfusion bottle 2 ventilate oxygen, make the oxygen content of culture medium reach 40%;

[0042] S6. Keep for 5 days, and keep the perfusion rate at 0.3RV / day;

[0043] S7, discharge, in the time of 8 days, begin to discharge, and keep its rate to be 0.1RV / day;

[0044] S8. The discharge is completed, and the stem cells are taken out from the perfusion box 5 to complete the stem cell culture. ...

Embodiment 2

[0046] A perfusion method for stem cell culture, specifically as follows:

[0047] S1, preparation work, prepare the stem cell culture medium, and lower the temperature of the stem cell culture medium to 10.5°C;

[0048] S2, put the stem cells into the container, and place it in the perfusion box 5;

[0049] S3, in the perfusion stage, connect the stem cell culture medium to the feed pipe 3, and enter the perfusion box 5 through the discharge pipe 4;

[0050] S4, turn on the peristaltic pump in the perfusion box 5, and keep the temperature at 33°C;

[0051] S5, ventilating oxygen, giving the perfusion bottle 2 ventilating oxygen, so that the oxygen content of the culture medium reaches 45%;

[0052] S6, keeping for 6 days, keeping the perfusion rate at 0.95RV / day;

[0053] S7, discharge, when 9 days, begin to discharge, and keep its rate to be 0.55RV / day;

[0054] S8. The discharge is completed, and the stem cells are taken out from the perfusion box 5 to complete the ste...

Embodiment 3

[0056] A perfusion method for stem cell culture, specifically as follows:

[0057] S1, preparation work, prepare the stem cell culture medium, and lower the temperature of the stem cell culture medium to 15°C;

[0058] S2, put the stem cells into the container, and place it in the perfusion box 5;

[0059] S3, in the perfusion stage, connect the stem cell culture medium to the feed pipe 3, and enter the perfusion box 5 through the discharge pipe 4;

[0060] S4, turn on the peristaltic pump in the perfusion box 5, and keep the temperature at 36°C;

[0061] S5, ventilating oxygen, giving the perfusion bottle 2 ventilating oxygen, so that the oxygen content of the culture medium reaches 50%;

[0062] S6, keeping for 7 days, keeping the perfusion rate at 1.6RV / day;

[0063] S7, discharge, when 10 days, start discharge, and keep its rate to be 1RV / day;

[0064] S8. The discharge is completed, and the stem cells are taken out from the perfusion box 5 to complete the stem cell c...

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Abstract

The invention relates to the technical field of stem cells, and discloses a perfusion device for embryonic stem cell culture. The perfusion device comprises a support, a flow limiting assembly and an extrusion assembly, the top end of the support is fixedly provided with a perfusion bottle, the inner wall of the perfusion bottle is fixedly provided with a feed pipe, and the inner wall of the perfusion bottle is fixedly provided with a discharge pipe. According to the perfusion device and the perfusion method for embryonic stem cell culture, a second electromagnetic block and a bottom shell are used in cooperation, in the ascending process of an electric telescopic column, a first electromagnetic block can be driven to ascend, the first electromagnetic block ascends to drive the second electromagnetic block to ascend, so that the second electromagnetic block drives a connecting rod to ascend, the bottom shell ascends, the bottom shell ascends to extrude a swing rod, so that the swing rod drives a first inner ring to rotate, a bent plate is driven to rotate, a guide rod is affected by the bent plate and extrudes the discharge pipe, the diameter of the discharge pipe is narrowed, the discharge amount of the discharge pipe is limited, and the flow of a culture medium is controlled.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to a perfusion device and a perfusion method for culturing embryonic stem cells. Background technique [0002] Stem cell research provides protection for people's health. Stem cells, as ancestor cells, can differentiate into various cells in the human body. This has a strong therapeutic guarantee for cancer or various serious diseases, but stem cells need to be extracted. When cultivating and treating cancer or serious diseases, a large amount of stem cells is required. If stem cells are not cultivated, the consumption of stem cells will be much higher than the extraction amount, resulting in a shortage of supply. [0003] At present, in the process of stem cell culture, it is generally carried out in the form of perfusion. Perfusion is to place the stem cells in the culture container, and continuously supply the medium through the connecting tube. The used medium is taken out o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/04C12N5/0735
CPCC12M23/48C12M29/06C12M29/12C12N5/0606
Inventor 姜胜维
Owner 姜胜维