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Compositions, devices and methods for the control in vitro of chemical microambient in cell cultures

A technology for in vitro cell culture, cell culture, applied in the field of compositions containing matrix, can solve the problems of expensive, difficult to manage, not repeatable, etc.

Pending Publication Date: 2019-07-19
ALMA MATER STUDIORUM UNIV DI BOLOGNA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method requires an expensive and difficult to manage setup with mechanical components that clearly cannot be replicated at the level of culture plates typically used for cell growth
Micrometric examination of concentrations and gradients of metabolites and soluble molecules in the extracellular matrix does not adequately reproduce the physiological environment

Method used

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  • Compositions, devices and methods for the control in vitro of chemical microambient in cell cultures
  • Compositions, devices and methods for the control in vitro of chemical microambient in cell cultures
  • Compositions, devices and methods for the control in vitro of chemical microambient in cell cultures

Examples

Experimental program
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Embodiment

[0061] 1.1 Hydrogel containing GOx and CAT

[0062] Glucose oxidase-GOx- is an enzyme of the oxidoreductase family, which catalyzes the following reactions:

[0063]

[0064] Hydrogen peroxide is a highly reactive molecule that is toxic to cells. For this reason, the above reaction is coupled with another very important oxidoreductase in the biological system-catalase -CAT-, which converts ROS substances produced in water. The net value of the two reactions is to convert two oxygen molecules into two molecules of water and oxygen, and the oxygen molecules will return to the glucose oxidase cycle. Hydrogel consumes oxygen in the presence of glucose, which is the basic substrate of cell metabolism in the cell culture device. These enzymes are proteins, and as such they have different reactive functional groups, such as amino acid groups ((-NH2) and carboxyl groups (-COOH)), they can be easily covalently bound to suitable reagents such as glutaraldehyde ( figure 1 ). Glutaraldehyde...

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Abstract

The present invention relates to compositions comprising a polymeric matrix or a gel containing functional enzymes capable of re-creating under culture conditions the cell microenvironment existing invivo. The present invention also relates to devices for cell cultures comprising such compositions, in particular hydrogel and the use thereof to control the chemical microenvironment of a cell culture or mimic physiological or pathological conditions of the in vivo cells. The compositions and the devices described herein could be also used in vitro for evaluating the therapeutic effect of a compound on a determined cell line or on primary cells.

Description

Technical field [0001] The present invention relates to a composition comprising a matrix containing functional enzymes capable of recreating the microenvironment of cells present in the body in a culture environment. The present invention also relates to a device for cell culture containing such a composition (especially a hydrogel composition), as well as a device for regulating the chemical microenvironment of cell culture or simulating the physiological or pathological environment of cells in vivo use. The composition and the device described herein can also be used to evaluate the therapeutic effect of a compound on defined cell lines or primary cells in vitro. Background technique [0002] Oxygen level and mechanical properties are the basic parameters of cell growth. These parameters regulate all functions and decisions belonging to the cell. Such properties need to be perfectly adjusted in in vitro cell culture. Generally, 20% oxygen is used for in vitro cell culture. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/09C12N11/02C12N11/06
CPCC12N5/0068C12N9/00C12N9/0006C12N9/0036C12N9/0053C12N9/0061C12N9/0069C12N9/0093C12Y101/03004C12Y106/03001C12Y109/03001C12Y110/03002C12Y111/01006C12Y113/12004C12Y117/03002C12N11/04C12N11/06C12N11/12C12N2500/02C12N2501/71C12N2533/50C12N2533/70C12N2537/10C12N11/087C12N11/089C12N5/0018C12N5/0693C12N9/0065C12N2533/30C12N2533/54C12N2533/72C12N2533/74C12N2533/76C12N2533/78C12N2533/80G01N33/5008
Inventor 斯特凡尼娅·拉皮诺弗朗切斯科·泽贝托盖斯顿·卡斯特拉尼斯蒂法诺·萨尔维奥利比阿特丽斯·法博尼伊莎贝拉·兹罗尼玛丽亚·孔特克劳迪·弗兰切斯基丹妮拉·萨尔瓦多麦拉·白克妮
Owner ALMA MATER STUDIORUM UNIV DI BOLOGNA
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