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Embryonic neural stem cell in-vitro culture medium and culture method

A neural stem cell and in vitro culture technology, applied in the field of embryonic neural stem cell in vitro medium and culture, can solve the problems of low purification efficiency, inability to in vitro culture of embryonic neural stem cells, and neural stem cell culture, etc.

Pending Publication Date: 2022-01-11
山东万海生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] For this reason, the present invention provides a kind of in vitro culture medium and culture method of embryonic neural stem cells, to solve the problem of neural stem cell culture, low purification efficiency and inability to culture embryonic neural stem cells well in the prior art due to simple rotational speed centrifugation. question

Method used

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  • Embryonic neural stem cell in-vitro culture medium and culture method
  • Embryonic neural stem cell in-vitro culture medium and culture method
  • Embryonic neural stem cell in-vitro culture medium and culture method

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Embodiment 1

[0029] This embodiment provides an in vitro culture medium for embryonic neural stem cells, including serum-free medium, EGF and bFGF. In the in vitro culture medium for embryonic neural stem cells, the concentration of EGF is 15-25ng, and the concentration of bFGF is 5-15ng.

[0030] The in vitro culture medium for embryonic neural stem cells in this embodiment includes serum-free medium, EGF and bFGF, wherein the serum-free medium can avoid the immune response caused by serum proteins; recombinant factors (EGF and bFGF) can stimulate the proliferation of neural stem cells, and the proliferation process In the medium, the neural stem cells grow on the wall, which can better remove other impurities or other cells, and the obtained neural stem cells are more pure.

[0031] The embryonic neural stem cell in vitro culture medium consisting of serum-free medium, EGF and bFGF of the present embodiment limits the concentration of EGF to 15-25ng, and the concentration of bFGF is 5-15n...

Embodiment 2

[0033] In the in vitro culture medium for embryonic neural stem cells as described in Example 1, the serum-free medium is DMEM / F12.

[0034]Serum-free media and reagents are widely used to culture mammalian and invertebrate cells to produce monoclonal antibodies, viral antigens and recombinant proteins, etc. Most serum-free media contain transferrin that transports ions into cells and insulin that regulates glucose uptake, as well as some proteins and albumin, fibrin, fetuin, etc. These proteins play various roles in cell culture. Functions, such as providing the matrix required for cell attachment, resisting the shear force of the bioreactor, and serving as a carrier of lipids and other growth and differentiation factors.

[0035] There are many kinds of serum-free medium, including UltraCULTURETM medium, PC-1TM medium, UltraMEM Reduced Serum medium, UltraCHOTM medium and so on. In the in vitro culture medium for embryonic neural stem cells in this example, DMEM / F12 was used...

Embodiment 3

[0037] In the in vitro culture medium for embryonic neural stem cells as described in Example 1 or 2, in the in vitro culture medium for embryonic neural stem cells, the concentration of EGF is 20 ng, and the concentration of bFGF is 10 ng.

[0038] The in vitro culture medium for embryonic neural stem cells of this embodiment is a preferred in vitro culture medium for embryonic neural stem cells, especially when the culture medium composed of DMEM / F12, EGF (concentration is 20 ng) and bFGF (10 ng) is used for in vitro culture of embryonic neural stem cells A better cultivation effect can be obtained. Under the concentration range defined in this example, EGF, bFGF and DMEM / F12 can play a good synergistic effect on each other.

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Abstract

The invention discloses an embryonic neural stem cell in-vitro culture medium and culture method. The embryonic neural stem cell in-vitro culture medium comprises a serum-free culture medium, EGF and bFGF. In the embryonic neural stem cell in-vitro culture medium, the concentration of the EGF is 15-25 ng, and the concentration of the bFGF is 5-15 ng. The embryonic neural stem cell in-vitro culture method comprises the steps: S1, inoculating embryonic neural stem cells into the embryonic neural stem cell in-vitro culture medium, putting the embryonic neural stem cell in-vitro culture medium into a CO2 cell culture box for culturing, and putting the cultured embryonic neural stem cells into a T25 culture bottle at the concentration of 5*105 pieces per 5ml; and S2, supplementing liquid into the T25 culture bottle at intervals of each 2-3 days, and after shaking, continuously culturing. According to the embryonic neural stem cell in-vitro culture medium and culture method, the neural stem cells are cultured by using a cell stimulating factor and the serum-free culture medium, so that the neural stem cells can be well subjected to in-vitro amplification and purification.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to an in vitro culture medium and culture method for embryonic neural stem cells. Background technique [0002] Neural stem cells are adult stem cells derived from the central nervous system, with self-renewal ability throughout life; they are also pluripotent stem cells, which can be induced to differentiate into various types of mature nerve cells, such as neurons, astrocytes and oligodendrocytes cell. Both the developing and mature central nervous system are distributed with neural stem cells. Neural stem cells are in the stage of rapid growth and development during the embryonic period. During this period, the neural stem cells mainly include cerebral cortex, striatum, hippocampus, olfactory bulb, cerebellum, spinal cord and other regions. There are still neural stem cells with the ability to proliferate and differentiate in the lateral ventricle wall and hippocampus of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797
CPCC12N5/0623C12N2500/90C12N2501/11C12N2501/115C12N2509/00C12N2509/10
Inventor 李相鲁赵克英
Owner 山东万海生物科技有限公司
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