Separation and purification method for spongy archaecyte

A technology for separation and purification of protocells, applied in the field of separation and purification of spongy protocells, which can solve the problems of high cell enrichment and achieve good separation effects, improved purity, and mild treatment methods

Inactive Publication Date: 2006-08-09
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, most of the sponge cell culture work at home and abroad uses mixed sponge cell culture. Even if the cell separation methods used are mostly density gradient centrifugation methods, they have not been able to highly enrich certain types of cells.

Method used

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  • Separation and purification method for spongy archaecyte
  • Separation and purification method for spongy archaecyte
  • Separation and purification method for spongy archaecyte

Examples

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Embodiment Construction

[0026] Isolation and Purification of Membrane Sponge Protocells in Huangbohai, China

[0027] Dissociation of Sponge Cells - Preparation of Hybrid Sponge Cells

[0028] Wash 2-3g of healthy luxuriant membranous sponge tissue blocks freshly collected or kept in an aquarium for 1-2 weeks with calcium-magnesium-free seawater (CMFSW) containing gentamicin (400ug / ml) for 3 times to remove the surface After microorganisms and other commensal organisms, cut into 1-3mm 3 Small pieces, stored at 4°C, washed 3-5 times with CMFSW, 2-3 hours each time, oscillating at a speed of 160rpm to separate out the sponge cells, filtered through a 300-mesh nylon filter, centrifuged at 1500rpm for 10min, and washed with 2mM EDTA CMFSW resuspended the cells to obtain mixed sponge cells, counted the percentage of protocells in the mixed cells with a hemocytometer, took samples to check the positive rate of PCNA, and then incubated some samples with BrdU at 37°C for 24 hours to check the mixed cells T...

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Abstract

This invention relates to a separation and purification method for sponge dry cells-original cells, which utilizes four methods of differential centrifugation, specific agglutination of cells, differential adherence and centrifugation of density and grads together to increase the purity and efficiency of separation and purification of sponge original cells and provides an effective method for studying the biological property of exuberate film sponge dry cells and the vitro culture rule.

Description

technical field [0001] The invention relates to the separation and purification of spongy protocells (stem cells), in particular to a method for isolating and purifying spongy protocells, which improves the purity and efficiency of separating protocells from mixed sponge cells. Background technique [0002] A large number of anti-tumor, anti-infection, and anti-HIV active substances with novel structure and strong activity can be isolated and extracted from sponge tissue. In order to obtain the above-mentioned active products, the research on sponge cell engineering based on sponge cell culture in vitro is also increasingly attracting attention. Attention, and become one of the most promising ways to solve the bottleneck problem of insufficient supply of sponge active substances. Sponges, the phylum Porezoa, are a class of low-level marine multicellular animals that feed on filtered seawater and have a rich ditch system, especially some smaller intertidal sponges, such as lu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/07
CPCC12N2500/25C12N2500/44C12N5/0601C12N2501/39C12N2500/36A61P31/18A61P35/00
Inventor 张卫孙黎明虞星炬金美芳赵权宇
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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