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Stem cell preparation as well as preparation method and application thereof

A kind of stem cell preparation and stem cell technology, applied in animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the problems of high recurrence rate, high long-term blindness rate, and inability to achieve a radical cure, so as to enhance resistance, Shorten the healing period, the effect of good curative effect

Inactive Publication Date: 2017-06-23
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, traditional Chinese medicine or clinical surgery are often used in the treatment of macular degeneration, but these methods can only delay the disease, but cannot cure it
In addition, existing treatments have a high rate of relapse and long-term blindness

Method used

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  • Stem cell preparation as well as preparation method and application thereof
  • Stem cell preparation as well as preparation method and application thereof
  • Stem cell preparation as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation of bone marrow mesenchymal stem cells

[0030] 1. Primary culture of bone marrow mesenchymal stem cells

[0031] Take the collected bone marrow and use ficoll density gradient centrifugation method in an ultra-clean bench to separate bone marrow mesenchymal stem cells; resuspend with DMEM+10% FBS complete medium, inoculate into a 15cm plate, and transfer to 5% CO 2 , Cultivated in an environment of 37°C.

[0032] After 48 hours of culture, the medium was changed, and new complete medium was added to continue the culture to obtain primary cells that could be subcultured; 2. Subculture of bone marrow mesenchymal stem cells

[0033] 1) Take the passable primary (P0) cells, discard the original culture medium in the dish, add 10mL PBS buffer solution to gently wash the cell growth surface, and discard the washing solution;

[0034] 2) Add 2mL of 0.25% trypsin, shake the plate left and right so that the trypsin evenly covers the bottom of the plate, u...

Embodiment 2

[0041] Preparation of embodiment 2 stem cell preparation

[0042] Mix albumin with physiological saline to prepare albumin suspension with a mass concentration of 5%;

[0043] Add EGF and TGF-β to the prepared albumin suspension, and adjust the concentration to 10 ng / mL;

[0044] Collect bone marrow mesenchymal stem cells of any generation from the second generation to the fifth generation prepared in Example 1, wash them twice with normal saline, resuspend the collected bone marrow mesenchymal stem cells with the suspension prepared above, and mix the bone marrow mesenchymal stem cells. The content of mesenchymal stem cells was adjusted to 3×10 6 cells, bone marrow mesenchymal stem cell preparations were obtained and stored in a 4°C environment for future use.

Embodiment 3

[0045] Preparation of embodiment 3 stem cell preparation

[0046] Mix albumin with physiological saline to prepare albumin suspension with a mass concentration of 5%;

[0047] Add EGF and TGF-β to the prepared albumin suspension, and adjust the concentration to 5 ng / mL;

[0048] The second to fifth generation bone marrow mesenchymal stem cells prepared in Example 1 were collected, washed twice with normal saline, and the collected bone marrow mesenchymal stem cells were resuspended with the albumin suspension prepared above, and the bone marrow mesenchymal stem cells were The content of mesenchymal stem cells was adjusted to 1×10 6 cells, bone marrow mesenchymal stem cell preparations were obtained and stored in a 4°C environment for future use.

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Abstract

The invention relates to the field of biotherapy, and discloses a stem cell preparation as well as a preparation method and application thereof. The stem cell preparation has a very good curative effect on macular degeneration. The stem cell preparation disclosed by the invention is prepared from the following components: 1*10<6> / ml to 5*10<6> of bone mesenchymal stem cells, 5ng / mL to 30ng / mL of epidermal growth factors, 5ng / mL to 30ng / mL of transforming growth factors-beta, albumin with the mass concentration of 5 percent, and a solvent. The invention further discloses the preparation method of the stem cell preparation; the preparation method comprises the following steps: a1), mixing the albumin and the solvent to prepare an albumin suspension solution; b1), adding the epidermal growth factors and the transforming growth factors-beta into the albumin suspension solution prepared by the step a1); c1), re-suspending the bone mesenchymal stem cells by utilizing a suspension solution prepared by the step b1), so as to prepare the stem cell preparation.

Description

technical field [0001] The invention relates to the field of biological therapy, in particular to a stem cell preparation and its preparation method and application. Background technique [0002] The macular area is an important area of ​​the retina, located at the posterior pole of the eye, and is mainly related to visual functions such as fine vision and color vision. Once lesions appear in the macular area, vision loss, black shadows in front of the eyes or visual distortion often occur. There are generally two types of macular degeneration. One is dry maculopathy and the other is wet maculopathy. Wet macular degeneration accounts for 10% of the total incidence. It is due to the abnormal growth of blood vessels under the retina, the new blood vessels rupture and bleed, and cause the growth of scar tissue, causing a sudden decrease in vision, which will quickly and seriously affect the central vision of the patient, and even cause Vision loss. The main symptoms of wet ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/38A61K38/18A61P27/02C12N5/0775A61K35/28
CPCA61K38/38A61K35/28A61K38/1808A61K38/1841C12N5/0663A61K2300/00
Inventor 葛啸虎陈海佳王一飞卢瑞珊王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD