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Particle-containing cell aggregate

A technology of aggregates and particles, applied in the field of cell aggregates, can solve the problems of reduced production capacity of useful substances and weakened cells

Inactive Publication Date: 2012-07-18
HITACHI HIGH-TECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in this case, as the size of the cell aggregate increases, the cells inside the aggregate become weaker and the ability to produce useful substances decreases.

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0039] A stirring propeller blade made of Teflon (registered trademark) was attached to a fixed stirring motor (manufactured by Shinto Scientific Co., Ltd., Sany type motor), and it was fixed together with a 1000 ml round bottom flask. 375ml of olive oil was added to the flask, and 10ml of an alkali-treated gelatin aqueous solution (concentration about 10%) with an isoelectric point of 4.9 was added dropwise while stirring at 37°C and 420rpm to prepare a W / O emulsion. After 10 minutes of stirring, the flask was cooled to 4°C and stirred for 30 minutes. After cooling, 100 ml of acetone was added thereto, stirred for 1 hour, and gelatin hydrogel particles were recovered by centrifugation. The recovered hydrogel particles were washed with acetone and further washed with 2-propanol to obtain uncrosslinked gelatin hydrogel particles. The hydrogel particles were dried and stored at 4°C.

[0040] 500 mg of dried uncrosslinked gelatin hydrogel particles were suspended in 100 ml of a...

Embodiment 2

[0044] Cell aggregates were produced in the same manner while changing the ratio of MSCs to the number of particles. As a result, when the number of particles / number of cells was 0.3 or more, the number of living cells increased with the culture time ( figure 2 ). That is, as the particle / cell ratio increases, the cell viability increases. This result is considered to be because the presence of the particles in the cell aggregate improves the diffusion of substances inside and outside the aggregate, thereby improving the nutrition of the cells, the supply of oxygen, and the excretion of waste.

Embodiment 3

[0046] Similar to Example 2, particle-containing MSC aggregates having a particle number / cell number ratio of 0.5 were produced, and the infiltration of BrdU into the aggregates was studied. On day 6 of cell culture, BrdU was added to the culture medium, and cell aggregates were collected on day 7 to make cryosections. Anti-BrdU antibodies were used to immunostain the largest section through the center of the aggregate. The stained sites indicated that the infiltration of BrdU into the nucleus was high and the cells were proliferating. The results confirmed the infiltration of BrdU into the cells on the surface and inside of the aggregate, indicating that the cells present inside were also proliferating ( image 3 ). In contrast, when cell aggregates were formed using MSC aggregates without particles and cultured under the same conditions, and the infiltration of BruU was studied, the infiltration was confirmed in cells near the surface of the aggregate, but not in the inner...

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Abstract

Disclosed is a particle-containing cell aggregate composed of cells and hydrogel particles, which are obtained by forming chemical crosslinks between one or more water-soluble synthetic polymers selected from among water-soluble synthetic polymers, polysaccharides and proteins. Also disclosed is a method for producing said cell aggregate.

Description

technical field [0001] The present invention relates to a particle-containing cell aggregate and a culture method thereof for improving the survival rate and biological function of cells. Background technique [0002] With the advancement of stem cell biomedical research, basic knowledge about cell differentiation and its biological function performance is constantly accumulating. Furthermore, by collecting adult (tissue) stem cells from various living body tissues, together with embryonic stem (ES) cells and iPS cells, research, drug development, and application of these cells to therapy become possible. Such development not only promotes the biomedical understanding of the proliferation, differentiation, and biological functions of stem cells, but also realizes the organization of cells by artificially controlling the differentiation and biological functions of cells. Drug development research and improving the effect of cell transplantation therapy are of great help. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/02C12N5/0775
CPCC12N2537/10C12N11/08C12N2533/54C12N5/0075C12N11/10C12N11/084C12N11/087
Inventor 田畑泰彦
Owner HITACHI HIGH-TECH CORP
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