Method for separating newly born rat olfactory bulb olfactory ensheat hing cells (OECs)

A technology of olfactory ensheathing cells and separation method, which is applied in the purification process of olfactory ensheathing cells and the field of separation of olfactory ensheathing cells of newborn rats, which can solve the problems of low purification efficiency and so on

Inactive Publication Date: 2012-07-11
吴卫江
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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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  • Method for separating newly born rat olfactory bulb olfactory ensheat hing cells (OECs)
  • Method for separating newly born rat olfactory bulb olfactory ensheat hing cells (OECs)
  • Method for separating newly born rat olfactory bulb olfactory ensheat hing cells (OECs)

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Embodiment Construction

[0014] 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:

[0015] 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.

[0016] The purification method of described neonatal rat olfactory bulb olfactory ensheathing cells (OECs) is as follows:

[0017] S1...

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Abstract

The invention provides a method for separating newly born rat olfactory bulb olfactory ensheat hing cells (OECs). The method is applied to the purification process of the OECs. The purification process comprises the following steps of: collecting the OECs, separating the OECs, and grouping, purifying and comparing the OECs; and performing morphologic observation and detecting the purity of the OECs. Through experiments of different methods for purifying the newly born rat olfactory bulb OECs, the newly born rat olfactory bulb OECs which are cultivated in vitro are mainly bipolar cells or three-pole cells, and the protuberances of the cells are thin and long. The OECs which are not purified are fibroblast which grows quickly, and is dominant; and the average value of the purity of the OECs which are inoculated after 14 days is more than 75 percent by the three purification methods. The purification rate of a p75 antibody adsorption method and a cytarabine inhibition method is slightly greater than that of a differential adhesion method. The cell purification of the newly born rat olfactory OECs is necessary in primary culture; and in research on spinal cord injury and regeneration, compared with the p75 antibody adsorption method and the cytarabine inhibition method, the differential adhesion method is simple, economic and practical method for purifying the OECs.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for isolating olfactory sheath cells of newborn rats, which is applied to the purification process of olfactory sheath cells. 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 there are not many research literatures on ...

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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