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Neural crest stem cell preparation and application thereof

A stem cell preparation and neural stem cell technology, applied in the field of stem cells, can solve the problems of limited self-renewal ability and achieve the effect of maintaining normal reproduction

Inactive Publication Date: 2018-11-06
广州沙艾生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A major feature of stem cells is their ability to generate multiple lineages and their ability to self-renew, whereas primary cultures of embryonic neural crest stem cells (NCSCs) have so far demonstrated self-renewal in vitro in contrast to bona fide stem cells ability is still limited

Method used

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preparation example Construction

[0038] A preparation method of the above-mentioned neural stem cell preparation, comprising the following steps:

[0039] S1. In vitro culture of neural tube and neural crest stem cells;

[0040] S2. Passage of neural crest stem cells;

[0041] S3. Proliferation of neural crest stem cells;

[0042] S4. Identification and evaluation of neural crest stem cell transplantation.

[0043] Preferably, the step S1 also includes the following steps:

[0044] (1) Coat the culture plate with Matrigel in advance, add 200ml of fresh serum-free medium to each well, put each neural tube into each well of a 96-well culture dish, at 37°C, the carbon dioxide concentration is 2%- Cultivate in 6% incubator;

[0045] (2) After 12 hours, fill each well with fresh medium to 1 mL, and gently tap the medium onto the wall of the culture dish with a pipette gun to avoid damaging the neural crest stem cells growing around the neural tube;

[0046] (3) After 12 hours, observe every two hours. When a ...

experiment example

[0065] Cell proliferation is the basis of biological reproduction and one of the important characteristics of cell life activities. Cell proliferation is used to maintain the relative balance of individual cell numbers, thereby maintaining their normal functions. In this experiment, the CCK8 assay method was used to further evaluate the proliferation ability of the third generation neural crest stem cells. The experimental results are shown in Table 1:

[0066] Table 1 Proliferation of neural crest stem cells

[0067] Days of cultivation (d)

[0068] Comprehensive analysis of the data in Table 1 shows that the neural crest stem cells grow gradually with the increase of the culture days. It shows that neural crest stem cells can grow stably in the medium we provide and maintain cell proliferation.

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Abstract

The invention specifically discloses a neural crest stem cell preparation and application thereof. The neural crest stem cell preparation provided by the invention is obtained by culturing a neural tube in vitro, wherein the neural tube is derived from a 10-day pregnant mouse embryo. The neural crest stem cell preparation disclosed by the invention and obtained by culturing the neural tube derivedfrom a 10-day pregnant mouse embryo in vitro identifies neural crest stem cells by immunofluorescence; P0 generation cells and P3 generation cells respectively express characteristics of neural crestand do not express the characteristics of neural stem, which indicates that the cultured cells successfully maintain the characteristics of mouse neural crest stem cells in vitro; and in-vitro culture experiments show that the neural crest stem cells can grow stably in a provided culture medium and normal proliferation of cells is maintained.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to a neural crest stem cell preparation and its application. Background technique [0002] Cells that are pluripotent and capable of self-renewal are called Embryonic stem cells (ESCs), which originate from the inner cell mass of the developing blastocyst, where pluripotency is the ability of a single cell to give rise to all the functions of a developing and adult organism. The ability of cell lineage; while self-renewal is the ability of cells to increase value in the same state. Pluripotency and self-renewal of embryonic stem cells (ESCs) is a major discovery for the control of molecular mechanisms. This feature makes it widely studied at different levels in the scientific field, from large-scale genomics and protein DNA research, to the study of the mechanism of individual regulatory factors [0003] The neural crest is a transitional structure in the development of vertebr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0735
CPCC12N5/0606C12N2500/05C12N2500/32C12N2501/20C12N2501/2302C12N2501/25
Inventor 徐智峰张新
Owner 广州沙艾生物科技有限公司
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