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Cryopreservation liquid and injection of mesenchymal stem cells

A technique for stem cells and cryopreservation, which is applied in the field of mesenchymal stem cell cryopreservation and injection, can solve the problems of high cost, difficulty in large-scale application, and difficulty in taking into account cell viability and direct clinical infusion, etc. The effect of simple method, low cost and good clinical application prospect

Active Publication Date: 2012-11-28
SICHUAN NEO LIFE STEM CELL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the cell cryopreservation solution contains human AB plasma, which contains a large number of unknown components, which poses a safety hazard when used for allogeneic transfusion. At the same time, there are few sources of plasma, and the cost is high, making it difficult to apply on a large scale
[0007] In summary, the cryopreservation solution of cells in the prior art is difficult to balance the long-term maintenance of cell viability and clinical direct infusion

Method used

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  • Cryopreservation liquid and injection of mesenchymal stem cells
  • Cryopreservation liquid and injection of mesenchymal stem cells
  • Cryopreservation liquid and injection of mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1 Preparation of Injection with Mesenchymal Stem Cell Freezing Solution of the present invention

[0079] 1. Preparation of mesenchymal stem cell injection of the present invention

[0080] As shown in Table 1, the formulation of the cryopreservation solution, take Bomaili A solution, 18AA 5% compound amino acid injection and 20% human serum albumin, mix well, add umbilical cord mesenchymal stem cells to a cell density of 1×10 7 pcs / ml, then add DMSO.

[0081] Table 1 Mesenchymal stem cell cryopreservation liquid formula of the present invention

[0082]

[0083] In group 6, the albumin content was high, and the cost was higher than other groups.

[0084] Control group: take 90ml of complete medium (90% (v / v) DMEM / F12+10% (v / v) fetal calf serum), mix well, add umbilical cord mesenchymal stem cells to a cell density of 1×10 7 pcs / ml, then add 10ml DMSO. The formula of the control group is a commonly used cell cryopreservation solution in China.

[0085] 2...

Embodiment 2

[0096] Example 2 Detection of various properties of the mesenchymal stem cell injection of the present invention after one year of frozen storage

[0097] 1. Experimental method

[0098] The mesenchymal stem cell injection of the present invention: Take another batch of cells that are prepared in the same way as in Example 1, Group 3, freeze them for 1 year and revive them in a water bath at 37°C, and store them in a refrigerator at 2-8°C.

[0099] Control group: with embodiment 1.

[0100] (1) Recovery after 1 year of cryopreservation, detection of cell viability at different time points after recovery

[0101] Take the cells before freezing to test the cell viability; test the cell viability of the mesenchymal stem cell injection of the present invention and the cells of the control group at 0 hours, 24 hours, 48 ​​hours and 72 hours after recovery. The detection method was classical trypan blue staining.

[0102] (2) Comparison of growth curves

[0103] Take the cells b...

Embodiment 3

[0130] Example 3 The relationship between cell viability and freezing time of mesenchymal stem cell injection of the present invention

[0131] 1. Experimental method

[0132] The mesenchymal stem cell injection of the present invention: prepare the mesenchymal stem cell injection according to the proportion of the first group and 3 in the embodiment, and store it in a liquid nitrogen tank after program-controlled cooling.

[0133] Contrast: with embodiment 1.

[0134] After 1, 6, and 12 months of frozen storage, they were revived in a 37°C water bath, and the cell viability was tested respectively. The detection method was the classic trypan blue staining method.

[0135] 2. Experimental results

[0136] The experimental results are shown in Table 5:

[0137] Table 5 The relationship between cell viability and freezing time

[0138]

[0139] As can be seen from Table 5, after 1, 6, and 12 months of cryopreservation, the cell viability of the mesenchymal stem cell injec...

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Abstract

The invention discloses a mesenchymal stem cell cryopreservation liquid which contains the following components by volume percent: 25-75% of Plasmalyte A, 10-35% of 18AA 5% compound amino acid injection, 1-50% of 20% human serum albumin and 5-20% of DMSO (dimethyl sulfoxide). The invention also provides a mesenchymal stem cell injection and a preparation method thereof. The mesenchymal stem cell cryopreservation liquid can overcome the defects of the traditional cell cryopreservation liquid, can maintain the motility of the mesenchymal stem cells for a long time, can also be directly clinically infused, and has excellent application prospects and economic benefits.

Description

technical field [0001] The invention relates to a mesenchymal stem cell cryopreservation solution and an injection solution. Background technique [0002] Stem cells (SC) are a kind of pluripotent cells with self-renewing ability, which can differentiate into various functional cells under certain conditions. Stem cells are not fully differentiated and immature cells, which have the potential to regenerate various tissues and organs and the human body. They are called "universal cells" in the medical field. [0003] Mesenchymal stem cells (mesenchymal stem cells, MSC) are important members of the stem cell family, derived from the mesoderm and ectoderm in the early stages of development. Originally found in bone marrow, MSCs have attracted increasing attention because of their multi-lineage differentiation potential, hematopoietic support and promotion of stem cell implantation, immune regulation and self-replication. Under specific induction conditions in vivo or in vitro...

Claims

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Application Information

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IPC IPC(8): A01N1/02A61K35/44A61K47/42A61K9/10A61P37/06A61P37/02A61P3/10A61P19/02A61P17/00A61K35/28
Inventor 任莹莹田甜冯学蓉
Owner SICHUAN NEO LIFE STEM CELL BIOTECH
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