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Method and formula for dissolving cell-loading micro capsule under physiological condition

A physiological condition, microcapsule technology, applied in the field of biochemistry, can solve the problems of large loss of cell activity, low cell recovery rate, large quantitative analysis error, etc., and achieve the effects of easy industrialization, high cell survival rate, and mild reaction conditions.

Inactive Publication Date: 2004-02-11
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on microcapsule breaking technology only retrieved one research paper on the mechanical breaking of APA microcapsules by Chang TMS in Canada, that is, placing a platinum metal ring in a centrifuge tube containing APA microcapsules, and passing through a high-speed The centrifugal rotation drives the platinum metal ring to shear the microcapsules, causing the microcapsules to break and release the cells in the capsules
However, this method has the following problems: ①The microcapsule breaking rate is low; ②The mechanical shear force causes a large loss of cell activity; ③There are many residual fragments in the microcapsules, and the cells are easy to adhere to the fragments, resulting in difficulty in subsequent separation; ④Quantitative The analysis error is relatively large; ⑤The cell recovery rate is low; this method has not yet been patented
[0004] The significance of microcapsule breaking technology lies in: ①under the premise of ensuring the survival of cells in the capsule, the microcapsules can be completely broken or melted; ②the quantitative characterization and monitoring of the growth state of the cells in the microcapsules can be realized; ③the current in vitro culture of cells The technology cannot completely simulate the growth environment of cells in vivo, so many cell lines must be cultured and proliferated in animals (such as intraperitoneal injection)
The outstanding problem is that a large number of host cells (such as macrophages) are mixed in the recovered cultured cells, and it is difficult to separate and purify

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0019] Example 1: Mix 8 mL of capsule burst solution containing 5 mol / L disodium edetate, 35 mmol / L sodium citrate, and 20 mmol / L sodium bicarbonate with 1.0 mL of APA microcapsules encapsulated with Escherichia coli in a test tube , shake manually for 10s. The APA microcapsules were all melted, and observed under a microscope, no microcapsule fragments were found, and a large number of free Escherichia coli cells could be observed in the solution, and the bursting liquid had basically no effect on the activity of Escherichia coli cells.

example 2

[0020] Example 2: Combine 10mL of capsule burst solution containing 5mol / L disodium edetate, 20mmol / L sodium bicarbonate, 35mmol / L sodium citrate with 1.0mL encapsulated yeast GS115 (cell density 1.69×107 / mL) of ACA microcapsules were mixed and shaken manually for 10s. The ACA microcapsules were all melted, and observed under a microscope, no residual fragments of the microcapsules were seen, and a large number of free yeast cells could be observed in the solution. After bursting the capsule, the survival rate of the cells in the bursting fluid was determined by the dilution plate method to be 80% to 95%. This example shows that the capsule rupture fluid not only ruptures the capsule completely and rapidly, but also basically has no effect on the activity of yeast cells.

example 3

[0021] Example 3: After directly contacting 10 mL of cystic rupture fluid containing 5 mol / L disodium edetate, 20 mmol / L sodium bicarbonate, and 35 mmol / L sodium citrate with adherent mucinous epidermal carcinoma cells in culture flasks for 20 minutes , dump the cystic fluid, and add mucoepithelial carcinoma cell culture fluid to the culture bottle. After 12 hours of culture, the cells can continue to adhere to the wall. This example shows that the cystic fluid has basically no effect on the viability of animal cells.

[0022] Advantages of the present invention can be summarized as follows:

[0023] (1) High rate of capsule rupture, up to 100%;

[0024] (2) The capsule breaking speed is fast, and the capsule breaking can be completed within 30 seconds.

[0025] (3) The pH value of the cystic rupture fluid is close to neutral, basically has no effect on the cell viability, and the cell survival rate can reach 80-95%.

[0026] (4) After breaking the capsule, the microcapsul...

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PUM

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Abstract

The present invention relates to biochemical technology, and is chemical process and formula of dissolving cell-loading microcapsule under physiological condition. The formula consists of complexoneIII, sodium citrate and sodium bicarbonate, and the present invention has the features of simple formula, mild reaction condition, fast capsule breaking speed, high cell survival rate, high capsule breaking rate and no microcapsule residue. The microcapsule breaking process maintains the cell activity and can fulfill the requirement in biomedicine application. The technology is favorable to theaccurate quantitative analysis of microcapsulated cell, the cell activity preservation, the obtaining of high purity and high yield cell or gene engineering product and the development of microcapsule to cell transportation function. The microcapsule breaking process is suitable for use in industrial production.

Description

technical field [0001] The invention relates to biochemical technology, in particular to a method and formula for dissolving cell-loaded microcapsules under physiological conditions by using a chemical method. Background technique [0002] With the development of material science and biotechnology, microcapsules with semi-permeable membranes have been applied in cell transplantation, cell culture, enzyme immobilization, drug controlled release and so on. In cell culture and transplantation research, the two most important types of microencapsulation systems are polylysine-sodium alginate-polylysine microcapsules (APA microcapsules for short) and chitosan-sodium alginate-chitosan Sugar microcapsules (ACA microcapsules for short) have the characteristics of mild preparation conditions and excellent biocompatibility. [0003] At present, researches related to APA or ACA microcapsules focus on the preparation of microcapsules, the interaction between microcapsule environment an...

Claims

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Application Information

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IPC IPC(8): B01J13/20C12N5/00C12N11/02
Inventor 马小军薛伟明于炜婷刘袖洞
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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