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Culture medium for rapid expansion of neural stem cells

A neural stem cell and culture medium technology, applied in the field of neural stem cells, can solve the problems of poor quality of stem cells, easy death of stem cells, single nutritional components, etc., and achieve the effects of preventing cell damage, ensuring balance, and ensuring integrity

Inactive Publication Date: 2020-01-21
深圳科学之门生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The single nutrient composition in the existing culture medium brings an unfavorable living environment for the proliferation of neural stem cells, the number of proliferation in the same time is low, and the quality of the proliferated stem cells is poor, which is not conducive to later research or use
At the same time, the nutrients in the medium do not have the function of protecting cells, and the proliferated stem cells are prone to death, which also brings an unreasonable environment for the survival of other stem cells.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Take 1L of basal medium, the content of each component of the nutrient components is: glucose 6g / L, glutamine 3*10 - 3 mol / L, NaHCO 3 5*10 -3 mol / L, penicillin 60mg / ml, streptomycin 65mg / ml, insulin 2.8μg / ml, transferrin 130ng / ml, progesterone 8*10 -9 mol / L, putrescine 30*10 -6 mol / L, selenium 50*10 -9 mol / L.

[0027] The above medium is used for the preparation of neural stem cells, including the following steps:

[0028] 1. Place the culture medium in a sterile box and use CO 2 Equilibrate for 30-40 min, CO 2 The concentration is 5%-10%, and the temperature is 32-34°C;

[0029] 2. Place the neural stem cells in the medium for culture, raise the temperature in the sterile box by 0.5°C every 2 hours until the temperature rises to 36.5-37.5°C, and culture for 5-10 hours;

[0030] 3. Add the culture medium in step 1 again to the aseptic box, repeat step 2, repeat this many times, the number of times to add the culture medium is 3-5 times, and the content of the a...

Embodiment 2

[0033] Take 1L of basal medium, and the content of the various components of nutrients is: glucose 15g / L, glutamine 8*10 - 3 mol / L, NaHCO 3 12*10 -3 mol / L, penicillin 90mg / ml, streptomycin 85mg / ml, insulin 3.5μg / ml, transferrin 250ng / ml, progesterone 15*10 -9 mol / L, putrescine 50*10 -6 mol / L, selenium 60*10 -9 mol / L.

[0034] The above medium is used for the preparation of neural stem cells, including the following steps:

[0035] 1. Place the culture medium in a sterile box and use CO 2 Equilibrate for 30-40 min, CO 2 The concentration is 5%-10%, and the temperature is 32-34°C;

[0036] 2. Place the neural stem cells in the medium for culture, raise the temperature in the sterile box by 0.5°C every 2 hours until the temperature rises to 36.5-37.5°C, and culture for 5-10 hours;

[0037] 3. Add the culture medium in step 1 again to the aseptic box, repeat step 2, repeat this many times, the number of times to add the culture medium is 3-5 times, and the content of the ...

Embodiment 3

[0040] Take 1 L of basal medium, the content of the various components of the nutritional ingredients is: glucose 10g / L, glutamine 5*10 -3 mol / L, 10*10 -3 mol / L, penicillin 75mg / ml, streptomycin 75mg / ml, insulin 3.0μg / ml, transferrin 190ng / ml, progesterone 12*10 -9 mol / L, putrescine 40*10 -6 mol / L, selenium 55*10 -9 mol / L.

[0041] The above medium is used for the preparation of neural stem cells, comprising the following steps:

[0042] 1. Place the culture medium in a sterile box and use CO 2 Equilibrate for 30-40 min, CO 2 The concentration is 5%-10%, and the temperature is 32-34°C;

[0043] 2. Place the neural stem cells in the medium for culture, raise the temperature in the sterile box by 0.5°C every 2 hours until the temperature rises to 36.5-37.5°C, and culture for 5-10 hours;

[0044] 3. Add the culture medium in step 1 again to the aseptic box, repeat step 2, repeat this many times, the number of times to add the culture medium is 3-5 times, and the content of t...

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Abstract

The invention discloses a culture medium for rapid expansion of neural stem cells. The culture medium comprises: a basic culture medium and nutrient components, wherein the basic culture medium is mixed solution of DMEM and F12; and the nutrient components comprise: glucose, glutamine, NaHCO3(subscript), penicillin, streptomycin, insulin, transferrin, progesterone, putrescine and selenium. The culture medium of the invention contains the insulin and the selenium, wherein the insulin plays an important role in the survival and growth of cells, and provides sufficient nutrient substances for thesurvival and growth of the cells; meanwhile, the selenium is a cooperative factor for the generation of glutathione, contributes to the hydrolysis of peroxide and superoxide, can prevent the damage of the cells, provides favorable conditions for the survival and rapid proliferation of the cells, and ensures the integrity of the cells. In a use method of the culture medium, CO2(subscript) is usedto balance the culture medium, the culture medium is added repeatedly, thus ensuring the sufficiency of the nutrient substances in the culture medium; additionally, the content of the culture medium is reduced by half each time, thus saving resources.

Description

technical field [0001] The invention relates to the field of neural stem cells, in particular to a culture medium for rapid expansion of neural stem cells. Background technique [0002] Neural stem cells are present in the nervous system and have the potential to differentiate into neurons, astrocytes and oligodendrocytes, so that they can produce a large number of brain cell tissues and perform self-renewal, which is sufficient to provide a large number of brain cells. A cell population of tissue cells. Neural stem cells have a strong ability to divide, and they need to exist in the culture medium for a long time to grow when they are cultured. [0003] The single nutrient composition in the existing culture medium brings an unfavorable living environment for the proliferation of neural stem cells, the number of proliferation in the same time is low, and the quality of the proliferated stem cells is poor, which is not conducive to later research or use. At the same time, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797
CPCC12N5/0623C12N2500/25
Inventor 胡俊杰
Owner 深圳科学之门生物工程有限公司
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