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Microcarriers, matrices and scaffolds for culturing mammalian cells and methods for their manufacture

A technology of copolymers and polymers, applied in general culture methods, chemical instruments and methods, medical preparations containing active ingredients, etc., can solve the problems of limited application of water-soluble cationic copolymers, and achieve the minimum cost and maximum output effect of reducing pollution

Active Publication Date: 2021-01-26
深圳鼎升生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Due to their known high cytotoxicity, water-soluble cationic copolymers currently have very limited applications in the biomedical field

Method used

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  • Microcarriers, matrices and scaffolds for culturing mammalian cells and methods for their manufacture
  • Microcarriers, matrices and scaffolds for culturing mammalian cells and methods for their manufacture
  • Microcarriers, matrices and scaffolds for culturing mammalian cells and methods for their manufacture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] Soluble 2,3-dialdehyde cellulose (DAC) or selectively oxidized mucin (SOM) can be prepared by sodium periodate oxidation of cellulose. 10 g of cellulose (size: <100 nm, 20 μm, 50 μm, or fibers) or 5 g of mucin (type II or III) can be resuspended by 200 mL of deionized water. Then, 20 g of sodium periodate was added, and then the pH was adjusted to 3.0 with 6X HCl. Next, the composition was degassed and purged with nitrogen, and then allowed to react at pH 3 for 4 hours at 60° C. with stirring in the dark. The reaction was stopped by adding 10 mL of ethylene glycol. The product was dialyzed against deionized water for 3 days. Soluble DAC or SOM was collected as a supernatant by centrifugation at 40,000 xg for 30 minutes to remove insoluble DAC or SOM as a pellet. Then, the collected supernatant is lyophilized (optional).

Embodiment 2

[0116] In another example, (low-oxidized) insoluble 2,3-dialdehyde cellulose (DAC) or selectively oxidized mucin (SOM) can be prepared by sodium periodate oxidation of cellulose. 10 g of cellulose (size: <100 nm, 20 μm, 50 μm, or fibers) or 5 g of mucin (Type II or Type III) were resuspended with 200 mL of deionized water, and then 10 g of sodium periodate was added. Then, the pH was adjusted to 3.0 with 6X HCl, followed by degassing and purging with nitrogen. The composition was then reacted at pH 3 at 60° C. for 4 hours with stirring in the dark. The reaction can be stopped by adding 10 mL of ethylene glycol. The product was washed 3 times with 4 liters of deionized water for 1 hour each. Insoluble DAC or SOM was collected by centrifugation at 800 xg for 10 minutes. Then, the washed insoluble DAC or SOM solution was resuspended with DI water (deionized water). The washed insoluble DAC solution can then be lyophilized (optional).

Embodiment 3

[0118] Polysaccharide-polyamine copolymers and glycoprotein-polyamine copolymers can be synthesized by reacting DAC or SOM with branched polyethyleneimine (PEI) (MW 750K). The ratio of DAC / SOM to PEI is 50:1 to 1:10. With branched polyethyleneimine 10g (MW 750K, in H 2 50% by weight in O) was added to a 500 mL beaker. The pH of PEI was adjusted to 5.0 with 37% HCl. The pH of 100 ml of DAC or SOM solution containing appropriate amount of DAC or SOM was adjusted to 5.0 by adding 6X HCl. Solutions of PEI and DAC or SOM were incubated on ice for 10 minutes. Solutions containing 10 g of PEI and solutions containing appropriate amounts of DAC or SOM were mixed and incubated on ice at 1000 RPM for 5 minutes. The mixture was kept on ice without agitation until complete product formation. The mixture was incubated at 70°C for 60 minutes. Check the pH of the suspension every 10 min and adjust it to 8.5 with 5M sodium hydroxide solution. The product is forced through a screen to o...

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Abstract

Disclosed herein are microcarriers, matrices and scaffolds for growing mammalian cells, including copolymer particles and matrices composed of polysaccharide-polyamine copolymers. The copolymer particles and matrix have a pore size of at least 50 microns and allow the growth of mammalian cells both on the outer surfaces of the particles and matrix and within the particles and matrix. Also disclosed herein are methods of making such microcarriers, matrices, and scaffolds, as well as combinations. In addition, provided herein are methods of growing mammalian cells using such microcarriers, matrices and scaffolds and combinations.

Description

[0001] This application claims the benefit of US Provisional Application No. 62 / 136,241, filed March 20, 2015, which is hereby incorporated by reference in its entirety. technical field [0002] In general, the present disclosure relates to materials for culturing anchorage-dependent mammalian cells, and in particular to vectors for high-density growth of anchorage-dependent mammalian cells, Substrates and scaffolds, and methods for their manufacture. Background technique [0003] The culture of mammalian cells, especially human cells, is critical in life sciences, pharmaceutical and biotechnology research, as cultures of human cells can be used to determine cell functions and interactions and, for example, to manufacture therapeutic cells, large quantities of Proteins and pathogens for vaccine development. Human gene products are commonly produced using in vitro methods via bacterial, yeast and insect gene expression systems. However, the normal biological function, activ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/715A61K31/717A61K31/738B01J20/24
CPCC08B15/02C08B15/06C08H1/00C08B15/005C12N5/0075
Inventor J.W.米歇尔杨达志
Owner 深圳鼎升生物科技有限公司
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