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Digestive juice used for separating anchorage-dependent cells and a separation method of the cells

An adherent cell and separation method technology, which is applied to the separation of adherent cells' digestive juice and the field of separation, can solve the problems of large differences, cumbersome operations, low cell survival rate, etc., and achieves reduction of differences in experimental results and dissociation efficiency. High, easy-to-use effects

Inactive Publication Date: 2018-01-09
MIAOSHUN SHANGHAI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the digestive solution and its separation method for separating adherent cells still have the following disadvantages: the dissociation efficiency of adherent cells is low, the damage to cells is relatively large, the toxicity is high, and the survival rate of cells is low; Adherent cells, multiple cell passages will affect the viability of the cells; the operation is cumbersome, and the experimental results between different operators vary greatly

Method used

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  • Digestive juice used for separating anchorage-dependent cells and a separation method of the cells
  • Digestive juice used for separating anchorage-dependent cells and a separation method of the cells
  • Digestive juice used for separating anchorage-dependent cells and a separation method of the cells

Examples

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Effect test

Embodiment 1

[0022] A digestive solution for separating adherent cells, including component A, component B and component C; component A is a cell washing solution, component B is a cell dissociation solution, and component C is a cell suspension ; The component A includes the following raw materials in parts by weight: 5 parts of NaCl, 0.1 part of KCl, 0.1 part of glucose, KH 2 PO 4 0.01 parts, Na 2 HPO 4 0.02 parts; the component B includes the following raw materials in parts by weight: 0.01 parts of dispase, 0.01 parts of papain, 500 parts of NaCl, 20 parts of KCl, 20 parts of glucose, KH 2 PO 4 2 parts, Na 2 HPO 4 2 parts; the component C includes the following raw materials in parts by weight: 4 parts of NaCl, 0.1 part of KCl, CaCl 2 0.2 parts, MgCl 2 0.1-part, 1 part of 4-hydroxyethylpiperazineethanesulfonic acid, 0.5 part of glucose.

[0023] A method for separating the digestive fluid of adherent cells, comprising the following steps: (1) removing the cell culture med...

Embodiment 2

[0025] A digestive solution for separating adherent cells, including component A, component B and component C; component A is a cell washing solution, component B is a cell dissociation solution, and component C is a cell suspension ; The component A includes the following raw materials in parts by weight: 11 parts of NaCl, 1 part of KCl, 2 parts of glucose, KH 2 PO 4 0.1 parts, Na 2 HPO 4 0.05 parts; the component B includes the following raw materials in parts by weight: 0.08 parts of dispase, 0.08 parts of papain, 900 parts of NaCl, 70 parts of KCl, 200 parts of glucose, KH 2 PO 4 8 parts, Na 2 HPO 4 8 parts; the component C includes the following raw materials in parts by weight: 10 parts of NaCl, 1 part of KCl, CaCl 2 0.5 parts, MgCl 2 0.5 parts, 4-hydroxyethylpiperazineethanesulfonic acid 4 parts, glucose 2 parts.

[0026] A method for separating the digestive fluid of adherent cells, comprising the following steps: (1) removing the cell culture medium, was...

Embodiment 3

[0028] A digestive solution for separating adherent cells, including component A, component B and component C; component A is a cell washing solution, component B is a cell dissociation solution, and component C is a cell suspension ; The component A includes the following raw materials in parts by weight: 8 parts of NaCl, 0.4 part of KCl, 1 part of glucose, KH 2 PO 4 0.06 parts, Na 2 HPO 4 0.0475 parts; the component B includes the following raw materials in parts by weight: 0.01-0.05 parts of dispase, 0.02-0.05 parts of papain, 800 parts of NaCl, 40 parts of KCl, 100 parts of glucose, KH 2 PO 4 6 parts, Na 2 HPO 4 4.75 parts; Described component C comprises the raw material of following parts by weight: 8.8 parts of NaCl, 0.224 parts of KCl, CaCl 2 0.44 parts, MgCl 2 0.19 parts, 2.38 parts of 4-hydroxyethylpiperazineethanesulfonic acid, and 0.9 parts of glucose.

[0029]A method for separating the digestive fluid of adherent cells, comprising the following step...

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Abstract

Digestive juice used for separating anchorage-dependent cells is disclosed. The digestive juice includes a component A, a component B and a component C. The component A includes NaCl, KCl, glucose, KH2PO4 and Na2HPO4. The component B includes dispase, papain, NaCl, KCl, glucose, KH2PO4 and Na2HPO4. The component C includes NaCl, KCl, CaCl2, MgCl2, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acidand glucose. The anchorage-dependent cell detachment efficiency of the digestive juice is high, and the digestive juice has low damage on cells and low toxicity and allows the cell survival rate to be high. A plurality times of cell passage can be performed without influences on cell vitality. The digestive juice can be used for cell detachment experiments for anchorage-dependent cells cultured in a serum-free manner. The method is simple to operate, and experiment result differences among different operators are reduced.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a digestive solution for separating adherent cells and a separation method thereof. Background technique [0002] Cell culture refers to a culture technique in which cells are removed from tissues in the body to simulate the living environment in the body, and are grown and reproduced while maintaining their structure and function under aseptic conditions, appropriate temperature, pH and certain nutritional conditions. The first culture of cells taken out of the body is primary culture, which is the initial and necessary stage of cell culture. When the primary cultured cells grow to a certain period, they need to be transferred to another container to continue to grow due to the restriction of the group environment. This process is called passage or subculture. At present, the digestive solution and its separation method for separating adherent cells still have the followin...

Claims

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Application Information

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IPC IPC(8): C12N5/00
Inventor 张彬张金保陈燕
Owner MIAOSHUN SHANGHAI BIOTECH CO LTD
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