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Mesenchymal stem cell preservation solution, preservation method and application thereof

A mesenchymal stem cell and preservation method technology, applied in the field of mesenchymal stem cell preservation solution, can solve the problems of low cell survival rate and inability to fully develop stem cells

Inactive Publication Date: 2019-09-06
深圳市未来细胞生命科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The problem with cell freezing for long-term storage of cells is that the survival rate of cells upon thawing is very low and cannot fully function as stem cells (Ali EO et al, 2019 / Wang WZ et al, 2013)

Method used

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  • Mesenchymal stem cell preservation solution, preservation method and application thereof
  • Mesenchymal stem cell preservation solution, preservation method and application thereof
  • Mesenchymal stem cell preservation solution, preservation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Preparation of fat stem cells

[0035] (1) Fat is extracted from the human abdomen or thighs by liposuction. The extracted fat and saline were mixed at a ratio of 1:1 and centrifuged at 300g for 3 minutes to remove oil, blood and saline. Fresh pure fat is extracted three times. The collagenase solution mixed with type I collagenase (SIGMA) and physiological saline at a ratio of 1:1 was mixed with fat at a ratio of 1:1, and the enzyme reaction was carried out at 37°C for 30-60 minutes. In addition to type I collagenase that is usually used in collagenase solutions, human collagenase that is not harmful to humans can also be used, and in this case, the reaction time is not set to 30 to 60 minutes. In particular, human collagenase is harmless to the human body and non-toxic, so no neutralization process is required. After the enzymatic reaction, centrifuge at 2000-2500g for 5 minutes to separate the oil layer and stem cell layer. At this time, the stem cells were in th...

Embodiment 2

[0042] will be 5×10 6 The adipose stem cells prepared in Example 1 were suspended in 3 mL of preservation solution containing 0.1% sodium hyaluronate, 0.3% albumin and 99.6% saline, and stored in a 5 mL syringe, so that the air content in the syringe was 0mL and refrigerated at 4°C. The refrigerated preservation solution was shaken up and down 44 times / min using a rocker (Nomi, Taizhou, NYC-80) to prevent cell sedimentation. Cells were stained with trypan blue solution (SIGMA), live cells and dead cells were counted using a hemocytometer, and cell viability was calculated. Monitor the survival rate of stem cells from 0h to 60h, see Table 2. The calculation of the cell viability in the following examples and comparative examples is the same as in Example 2.

[0043] Table 2 The effect of preservation solution on the survival rate of stem cells at 0-60h

[0044]

[0045]

[0046] It can be seen from Table 2 that the survival rate of stem cells can reach 81.76% at 24 ho...

Embodiment 3

[0048] will be 5×10 6 The adipose stem cells prepared in Example 1 were suspended in 3 mL of preservation solution containing 0.3% sodium hyaluronate, 0.3% albumin and 99.4% saline, and stored in a 5 mL syringe, so that the air content in the syringe was 0mL and refrigerated at 4°C. The refrigerated preservation solution was shaken up and down 44 times / min using a rocker (Nomi, Taizhou, NYC-80) to prevent cell sedimentation. Monitor the survival rate of stem cells from 0h to 60h, see Table 3.

[0049] Table 3 Effect of Preservation Solution on Stem Cell Survival Rate at 0~60h

[0050]

[0051] It can be seen from Table 3 that the survival rate of stem cells can reach 86.78% at 24 hours and 60.42% at 48 hours using the preservation solution of Example 3.

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Abstract

The invention provides a mesenchymal stem cell preservation solution, a preservation method and application thereof, and relates to the technical field of stem cell preservation solutions. The mesenchymal stem cell preservation solution provided by the invention comprises the following components in percentage by volume: 0.1 to 0.5 percent of sodium hyaluronate, 0.2 to 0.4 percent of albumin and the balance of physiological saline. The survival rate of stem cells can reach more than 80% within 24 hours by using the preservation solution provided by the invention.

Description

technical field [0001] The invention relates to the technical field of stem cell preservation solution, in particular to a mesenchymal stem cell preservation solution and its preservation method and application. Background technique [0002] Mesenchymal stem cells can be isolated from various tissues, such as bone marrow, adipose tissue, skin, skeletal muscle, and cord blood. Among them, stem cells derived from adipose tissue are called adipose stem cells (ASC) (Gimble et al., 2011). Adipose-derived stem cells can be easily obtained with minimal surgical procedures, and the cell yield is higher than typical bone marrow-derived mesenchymal stem cells (MSC) (B.cousin et al., BBRC 301:1016,2003; A.Miranville et al., Circulation, 110:349, 2004; S. Gronthose et al., J. Cell Physiol. 189:54, 2001; M.J. Seo et al., BBRC, 328:258, 2005). It is considered to be the most useful mesenchymal stem cell because it is rich in growth factors and cytokines (Sgodda et al., 2007, P.A. Zuk et...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 李承根李松伊成大一张涛徐源
Owner 深圳市未来细胞生命科技有限公司
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