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Grouping purification method of olfactory ensheathing cells of neonatal rat

A purification method and technology of olfactory ensheathing cells, which can be applied to animal cells, nervous system cells, vertebrate cells, etc., can solve the problem of low purification efficiency.

Inactive Publication Date: 2012-07-25
吴卫江
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

There are many methods for purifying OECs cells, each with its own advantages and disadvantages. In the study of OECs cell transplantation to repair SCI, there is no conclusion about which purification method to use. At the same time, there are not many research literatures on the comparison of purification efficiency.

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  • Grouping purification method of olfactory ensheathing cells of neonatal rat
  • Grouping purification method of olfactory ensheathing cells of neonatal rat

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Embodiment Construction

[0013] Such as figure 1 A flow chart of the steps of an embodiment of the purification method of the newborn rat olfactory bulb olfactory ensheathing cells (OECs) of the present invention shown, the materials used in the described embodiment are as follows:

[0014] Neonatal 1-2d SD rats were purchased from the Experimental Animal Center of Soochow University School of Medicine; DM EM / F212 medium was purchased from In2vit Rogen Company, trypsin, Forskolin, bovine pituitary extract (B PE), cytarabine ( Ara c), polylysine were purchased from Sigma Company, and calf serum was purchased from Hangzhou Sijiqing Company. Glial fibrillary acidic protein GFA P antibody, low-affinity neurotrophic factor receptor p75 antibody, biotinylated goat anti-rabbit IgG, and SABC coupled with horseradish peroxidase (HRP) were purchased from Wuhan Boster Company.

[0015] The purification method of described neonatal rat olfactory bulb olfactory ensheathing cells (OECs) is as follows:

[0016] S1...

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Abstract

The invention provides a grouping purification method of olfactory ensheathing cells (OECs) of a neonatal rat, and the method is applied to a purification process of the OECs of the neonatal rat, wherein the purification process comprises the following steps of: obtaining the OECs; separating the OECs; performing grouping purification and comparison on the OECs; and performing morphologic observation and OECs purity detection. By virtue of different purification method experiments of the OECs of the neonatal rat, results indicate that the OECs of an in-vitro cultured neonatal rat are mainly bipolar or tripolar cells, and the neurite of the cell is fine. The OECs which are not purified have quickly-growing and dominated fibroblast, and the three purification methods realize that the purity average value of the OECs after being inoculated for 14 days is more than 75 percent. The purification rates of a p75 antibody adsorption method and a cytarabine inhibition method are slightly higher than that of a difference-speed adherence method. Therefore, cell purification in OECs primary culture of the neonatal rat is necessary; and in a spinal cord injury and regeneration research, compared with the p75 antibody adsorption method and the cytarabine inhibition method, the difference-speed adherence method is a simple, economical and practical OECs purification method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for grouping and purifying olfactory ensheathing cells of neonatal rats, which is applied to the purification process of olfactory ensheathing cells of neonatal rats. Background technique [0002] In recent years, olfactory ensheath hing cells (OECs) have become a hotspot in the field of spinal cord injury (SCI) repair theory and applied basic research, and are considered to have great application prospects in the research of tissue cell transplantation repair SCI. transplant material. As a transplant material, OECs have a large demand and high requirements for purity and activity, so they must be obtained through in vitro culture and purification. There are many methods for purifying OECs cells, each with its own advantages and disadvantages. In the study of OECs cell transplantation to repair SCI, there is still no conclusion about which purification method to use, and th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079
Inventor 吴卫江
Owner 吴卫江
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