Composition containing mesenchymal stem cell-hydrogel and method for producing the composition

A technology of stem cells and hydrogels, applied in biochemical equipment and methods, cell culture supports/coatings, medical preparations containing active ingredients, etc., can solve problems such as basement membrane protein damage and cell loss, and achieve The effect of preventing cell loss

Inactive Publication Date: 2017-08-18
ANTEROGEN
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned documents only disclose that when fibrin / HA is used as a support, it is possible to promote mesenchymal function without adding transforming growth factor β (TGF-beta) added for conventional cell differentiation. Mesenchymal stem cells differentiate into chondrocytes
[0007] In the case of previous methods, enzymatic treatment was performed during final cell preparation for transplantation, thus causing damage to intercellular junctions and basement membrane proteins, during cell collection, washing, vial filling steps, and refilling from vials to syringes. In each step, cell loss occurs, and there is a problem that after enzyme treatment, only single cells are collected and injected, and therefore, most active substances such as collagen synthesized during cell culture are removed

Method used

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  • Composition containing mesenchymal stem cell-hydrogel and method for producing the composition
  • Composition containing mesenchymal stem cell-hydrogel and method for producing the composition
  • Composition containing mesenchymal stem cell-hydrogel and method for producing the composition

Examples

Experimental program
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Effect test

Embodiment 1

[0050] Example 1. Human adipose-derived mesenchymal stem cell culture method

[0051] Usually, adipose tissue is obtained by liposuction, but not limited thereto.

[0052] Adipose-derived mesenchymal stem cells were isolated from adipose tissue obtained by liposuction by the following method. To remove blood, the adipose tissue was washed 3 to 4 times with phosphate-buffered saline (PBS) having the same volume as the adipose tissue. Put the same volume of collagenase solution as the volume of adipose tissue, and react in a water bath at 37°C. The adipose tissue was transferred to a centrifuge tube, and centrifuged at 20° C. and 1500 rpm for 10 minutes. Remove the fat layer as the supernatant, and carefully and smoothly separate the collagenase solution in the lower layer. After placing in the substrate for culture and suspending, centrifugation was performed at 20° C. and 1,200 rpm for 5 minutes. At this point, the matrix-vascular components are precipitated, and the super...

Embodiment 2

[0053] Example 2. Determination of the concentration of fibrin glue gel

[0054] To the 5×10 obtained in Example 1 above 5 Thrombin solution (400-600 I.U.) was added to the adipose-derived mesenchymal stem cells / mL at a ratio of 1:10, 1:20, and 1:40. Prepare by diluting fibrinogen (71.5-126.5 mg / mL) at ratios of 1:20, 1:40, and 1:80, respectively. 500-700 μL of the cell suspension containing fibrinogen and thrombin were added to each well of the 12-well plate using a double-barrel syringe system. If the gel is completely solidified, after adding a medium containing 10% fetal bovine serum and 1 ng / mL basic fibroblast growth factor, the 2 Cultivation was carried out in an incubator for 5 days. After washing with Dulbecco's modified Eagle's medium three times, add Dulbecco's modified Eagle's medium containing 1 ng / mL basic fibroblast growth factor, at 37°C, 5% CO 2 The cultivation was carried out for about 12 hours in the incubator. The supernatant was removed, and the cell-...

Embodiment 3

[0060] Example 3. Preparation of cell-hydrogel culture fluid

[0061] To the 5×10 obtained in Example 1 above 5 Adipose-derived mesenchymal stem cells / mL were prepared by adding thrombin solution at a ratio of 1:20, and diluting fibrinogen at a ratio of 1:40. Then, cultured according to Example 2, washed, and injected into the syringe filling.

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Abstract

The present invention relates to a mesenchymal stem cell composition cultured in hydrogel, and more specifically to a composition containing human fat cell-derived mesenchymal stem cell which is stable and in a form allowing immediate administration, and a method for producing the composition. More specifically, by providing a composition in a form allowing immediate administration by filling a syringe with the mesenchymal stem cells after culturing same in hydrogel and washing, an enzyme treatment in the final cell transplantation preparation process is obviated, and as washing is carried out in the hydrogel form without passing through a washing step utilizing centrifugation and the like, damage to the cells is nearly nonexistent, and as biologically active substances are contained within the hydrogel pores, the present invention has the benefit of a treatment effect immediately following the administration into an individual.

Description

technical field [0001] The present invention relates to a mesenchymal stem cell composition cultured in a hydrogel, in more detail, to a human adipose-derived mesenchymal stem cell-hydrogel composition and a preparation method thereof, more specifically, to a hydrogel cultured After the mesenchymal stem cells are washed and filled into the syringe, administration can be performed immediately. In the final preparation of the transplanted cells, there is no need for enzyme treatment, and there is no need for cleaning steps such as centrifugation. Instead, the hydrogel Therefore, there is no loss of cells, and the physiologically active substance is inserted into the pores of the hydrogel, thereby having the advantage of exhibiting a therapeutic effect after administration to an individual. Background technique [0002] In the process of protease treatment of isolated cells obtained by protease treatment such as trypsin or dispase, conventional first-generation stem cell therap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/12A61K9/06C12N5/0775C08J3/075
CPCA61K9/06A61K35/28A61K38/363A61K38/4833A61K38/014C12N2533/52C12N5/0667C12N2539/00C12N5/0068A61K2300/00A61K48/00C08J3/075A61K35/12A61K47/42C08J2389/00C12N2533/56
Inventor 李晟丘金美馨尹正仁
Owner ANTEROGEN
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