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Cell microcarrier and preparation method thereof

A technology of cell microcarriers and hydroxyapatite, which is applied in the direction of microcarriers, general culture methods, biochemical equipment and methods, etc. It can solve the problems of increasing the preparation cycle and cost of microcarriers, difficult cell adhesion, and the decline of mechanical properties of microcarriers, etc. problems, to achieve excellent biocompatibility and biological activity, good biocompatibility and degradability, and favorable effects of adhesion and proliferation

Inactive Publication Date: 2017-03-08
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Application Information

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Problems solved by technology

However, the use of conventional single polymer components, such as polymethyl methacrylate (PMMA), polylactic acid-glycolic acid copolymer (PLGA), etc., has poor cell compatibility, and cells are not easy to adhere to the surface; For example, adding hydroxyapatite can promote the adhesion and proliferation of bone cells, but it has poor compatibility with polymers, is not easy to disperse evenly in polymer components, and is brittle and lacks plasticity, which will lead to poor mechanical properties of microcarriers. Decrease or affect its spherical shape, etc. At the same time, there will be more pre-mixing steps, which increases the cycle and cost of microcarrier preparation

Method used

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  • Cell microcarrier and preparation method thereof
  • Cell microcarrier and preparation method thereof
  • Cell microcarrier and preparation method thereof

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preparation example Construction

[0028] The present invention also provides a preparation method of the above-mentioned cell microcarrier, comprising the following steps: S1) Stirring and reacting hydroxyapatite modified with a silane coupling agent and amino acid-N-carboxylic acid anhydride in an organic solvent to obtain grafted Polyamino acid hydroxyapatite; S2) mixing the grafted polyamino acid hydroxyapatite with a volatile organic solvent to obtain a mixed solution; mixing the mixed solution with an aqueous solution of a water-soluble polymer compound, stirring , removing volatile organic solvents, and washing with water to obtain cell microcarriers.

[0029] The present invention has no special limitation on the sources of all raw materials, which can be commercially available.

[0030] Wherein, the preparation method of the hydroxyapatite modified by the silane coupling agent is a preparation method well known to those skilled in the art, and there is no special limitation. In the present invention, t...

Embodiment 1

[0044] 1.1 Co-precipitation method to prepare hydroxyapatite modified by silane coupling agent KH-550 (referred to as HA-APS): add hydroxyapatite (HA) to 90% ethanol aqueous solution of silane coupling agent KH-550, The mass ratio of HA to silane coupling agent KH-550 was 1:0.2, stirred at room temperature for 6 hours, and then the product was washed with ethanol for 3 to 5 times, dried at room temperature, and treated at 130°C for 2 hours to obtain HA-APS.

[0045] 1.2 With anhydrous dioxane as solvent, HA-APS and benzyl glutamate-N-carboxylic acid anhydride (referred to as BLG-NCA) according to the mass ratio (1:1-1:2), after 32 hours of reaction, the preparation The polybenzyl glutamate grafted hydroxyapatite (referred to as PBLG-g-HA) with a grafting amount of 30%-50% (1:1 can prepare the PBLG-g-HA with a grafting amount of about 33%) , denoted as P-g-H-33; 1:2 can prepare PBLG-g-HA with a grafting amount of about 50%, denoted as P-g-H-50).

[0046] 1.3 Fully disperse PBL...

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Abstract

The invention provides a cell microcarrier and a preparation method thereof; the preparation method comprises the steps of stirring and reacting silane coupling agent-modified hydroxyapatite and amino acid-N-carboxylic anhydride in an organic solvent to obtain polyamino acid-grafted hydroxyapatite; mixing the polyamino acid-grafted hydroxyapatite with an easily-volatile organic solvent to obtain mixed solution; mixing and stirring the mixed solution with water-soluble polymer compound solution, removing the easily-volatile organic solvent, and washing with water to obtain the cell microcarrier. Compared with the prior art, the cell microcarrier and the preparation method thereof have the advantages that in-situ ring-opening polymerization of amino acid-N-carboxylic anhydride monomers is initiated through amino surface-modified hydroxyapatite, amino acids are polymerized in situ to the surface of hydroxyapatite, and the prepared cell microcarrier is free of other polyester materials, effectively combines the advantages of hydroxyapatite and polyamino acid, facilitates the adhesion and proliferation of cells, and is suitable for in-vitro cell amplification and in-vivo bone tissue repairing.

Description

technical field [0001] The invention belongs to the technical field of biological materials, and in particular relates to a cell microcarrier and a preparation method thereof. Background technique [0002] Microcarrier is a kind of tiny spherical particle, which can make adherent cells grow in a single layer on the surface of the particle in a suspended state in the three-dimensional cell culture, so it has a larger contact area than ordinary flat culture, which is conducive to the growth of cells. Scale cultivation and collection. The proliferation of adherent cells on the surface of microcarriers needs to go through the following three stages: contact and adhesion of cells to the surface of microcarriers, growth and expansion to monolayer cells. Among them, the contact and adhesion between cells and the surface of microcarriers is the most important link. Only when cells are well adhered to the surface of microcarriers can they grow and proliferate further. Whether cells...

Claims

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Application Information

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IPC IPC(8): C12N5/077C08L29/04C08K9/06C08K9/04C08K3/32
CPCC12N5/0068C08K3/32C08K9/06C08K9/08C08K2003/325C08L2201/06C08L2203/02C12N5/0654C12N2531/00C08L29/04
Inventor 章培标李林龙付川王宗良史新翠高田林
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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