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In-vitro schistosomulum co-culture method

A technology of co-culture and schistosomiasis, applied in the field of biomedicine, can solve problems such as differences in gene expression

Inactive Publication Date: 2011-01-26
WUHAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, studies have shown (Chai M, McManus DP, McInnes R, et al.Cell Mol.Life Sci., 2006, 63(7-8):919-929), the larvae of Schistosoma japonicum cultured by conventional in vitro culture methods, although In terms of ultrastructure, it is not much different from the larvae in the final host, but there are significant differences in gene expression
Therefore, it is considered that it is necessary to be cautious when the larvae of Schistosoma japonicum cultured by conventional in vitro culture methods replace the larvae in the host for molecular biology and immunology research

Method used

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Examples

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Embodiment 1

[0029] Example 1: Comparison between co-cultured larvae for 3 days and larvae in the host, and routine culture in vitro for 3 days

[0030] 1. Establishment of co-cultivation method of Schistosoma japonicum larvae and host cells ( figure 1 )

[0031](a) Take the positive snails and release the cercariae by the conventional method, paste the escaped cercariae with a sterilized cover glass with a size of 12×25mm, and put 10ml of cercariae washing solution (containing 0.5% hydrolyzed milk protein, 300IU / ml penicillin and 300ug / ml streptomycin (Earle's solution) at 4°C for 10-15min, centrifuged at 700g for 3min, removed the supernatant and coverslip, washed with cercariae washing solution, and centrifuged aseptically for 5-6 times. Then add 4-5ml of washing solution and mix with the cercariae, suck the cercariae suspension into a sterilized 10ml syringe, connect it to another sterile 10ml syringe through a sterile conjoined needle, and push the cercariae repeatedly and quickly ...

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Abstract

The invention relates to the field of biomedicine and discloses a schistosomulum co-culture method by co-culturing schistosomulum and definitive host cells thereof. Proved by experiments, Japanese schistosomulum cultured for 3 days by using the co-culture method of the invention not only has the surface structure, the tegument structure, and the like more similar to a 3-day lung-stage schistosomula growing in a host than a schistosomula cultured for 3 days by using a conventional in-vitro culture method, but also has the gene expression and the soluble proteins characteristics more approaching to the schistosomula growing in the host. The culture method of the invention more approaches to an in-vitro schistosome culture method of in the in-vivo environment of the host, is beneficial to the research on schistosome genomics, post-genomics, transgenic and proteomics and schistosomiasis immunology and lays a foundation for disclosing a growth and development mechanism of the schistosome and the mutual relation between the schistosome and the host.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the establishment of a new in vitro culture method for artificially transforming Schistosoma japonicum larvae and their final host cells. Background technique [0002] The pulmonary stage of schistosomiasis in the final host is regarded as the best target for host immune system attack, and its antigen can induce Th1-mediated protective immune response. Therefore, the specific target molecule of larvae is considered to be the best candidate molecule for the new generation vaccine of schistosomiasis, and its screening is extremely important for the development of schistosomiasis vaccine. However, the material source of lung-stage larvae is relatively difficult, and the percentage obtained from the infected final host is only 30%, which affects the extraction of a large number of target antigens and the understanding of their characteristics. At present, domestic and foreign research nee...

Claims

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Application Information

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IPC IPC(8): A01K67/033
Inventor 董惠芬叶青赵琴平明珍平钟沁萍朱俊勇李瑛蒋明森
Owner WUHAN UNIV
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