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Culture process of human nerve stem cell

A culturing method and stem cell technology, applied in the field of culturing human neural stem cells, can solve the problem of not being able to seed cells to cultivate neural stem cells, and achieve the effects of reducing the number of fibroblasts, having good effects and reducing toxicity

Inactive Publication Date: 2004-10-20
徐如祥 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main function of the existing synthetic cell culture medium is to provide a general survival and growth microenvironment for different types of tissue cells, such as Eagle, DMEM, F12, RPMI1640, CMRL1066, L15, 199, MB752 / 1, etc., which mainly contain components of Amino acids, vitamins, inorganic salts, sugars, etc., the method of cultivating cells using these synthetic media cannot cultivate seed cells from different sources (such as bone marrow tissue cells, etc.) into the required neural stem cells

Method used

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Embodiment Construction

[0018] This embodiment includes the following steps:

[0019] 1. Prepare human neural stem cell medium. The preparation steps of this medium are: a. Take 12g of basal culture medium mixed with DMEM and F12 in a ratio of 1:1, add pure water to a concentration of 12g / L, and fully stir to dissolve , make the general basal culture medium of cell; b, take again insulin (Insulin) 5mg, L-glutamine (L-Glutamine) 3.9mg, butanediamine hydrochloride (Putrescine) 10mg, sodium selenide (Sodium Selenide) 0.03 mg, human transferrin (Transferrin) 50mg, hydrocortisone (Hydrocortisone) 10mg, progesterone (Progesterone) 0.02mg were sequentially added to the basal culture medium prepared in step 1, fully stirred evenly, and made up to volume with purified water It was 1000 milliliters, and now it was in an orange-yellow slightly turbid state; c, adjusting the pH to 7.8 with 5 normal concentrations of sodium hydroxide, now the culture medium was in a pink and clear state; d, the laminar flow cell ...

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Abstract

The invention discloses a cultivated method of nervous stem cells, it includes following steps:1, Nervous stem cells substrate of human being is made up of base cultivate liquid which is compounded by mixing DMEM and F12 by rate of 1:1, and insulin, muriatic acid butyl amic, selenium natrium, ydrogen cortisone, L-glutamic acyl amic, man-turn iron albumen, flavone; 2, Noumenon serum of patient themselves is gathered immediately; 3. Noumenon serum of patient themselves is put into compounded Nervous stem cells substrate of human being, them, it is incubated by adding 3-7% CO2 warmer under conditino of 35C-38C. The method of the invention is simple, and invention has better repetition and its operation is simple and convenient; it plays an inestimable part in researching, teaching, clinic application, etc all of which relate to nervous stem cell and its application.

Description

technical field [0001] The invention relates to a cell culture method, in particular to a culture method of human neural stem cells. Background technique [0002] Neural stem cell research is a new research topic at home and abroad since 1998. Neural stem cells are pluripotent cells of the central nervous system and common precursor cells of the three main cells in the brain (neurons, astrocytes, and oligodendrocytes), and have the following characteristics: (1) in an undifferentiated state (2) have multilineage differentiation potential, that is, the ability to evolve into different types of mature cells; (3) can self-replicate or renew through asymmetric division, and produce the same offspring as itself cells to maintain a stable cell number. There are four main sources of neural stem cells: embryonic stem cells or embryonic neural tissue, neural crest cells, brain tissue, and bone marrow stroma. Just because neural stem cells have the ability to differentiate into neu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/00C12N5/0797
Inventor 徐如祥姜晓丹
Owner 徐如祥
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