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Mesenchymal stem cell low-temperature preserving fluid and preparation method thereof

A technology of mesenchymal stem cells and cryopreservation, applied in the field of stem cell preservation, can solve the problems of inability to use, decreased cell viability, and cannot be used as a preservation solution for mesenchymal stem cells for clinical treatment, etc. Avoid the effects of the freeze-thaw step

Inactive Publication Date: 2017-07-07
浙江新生泉细胞科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

M199 and PBS are not clinically approved drugs for injection, so they cannot be used as a preservation solution for mesenchymal stem cells for clinical treatment
DMEM is a cell culture medium, but it is only used for scientific research, not as a reagent for clinical use
Although 0.9% normal saline and 5% glucose are clinically safe preservation solutions, the survival rate of these two preservation solutions decreases rapidly, and they are only suitable for immediate use of mesenchymal stem cells and cannot be used for a long time (such as 24h) use after transportation

Method used

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  • Mesenchymal stem cell low-temperature preserving fluid and preparation method thereof
  • Mesenchymal stem cell low-temperature preserving fluid and preparation method thereof
  • Mesenchymal stem cell low-temperature preserving fluid and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Comparison of cell viability, adherence and clone formation between the mesenchymal stem cell injection prepared from a mesenchymal stem cell cryopreservation solution of the present invention and injections prepared from other formulations

[0029] 1.1 Preparation of mesenchymal stem cell injection of the present invention and other formula injections

[0030] The formulations, storage temperature and storage time of different storage solutions are shown in Table 1.

[0031] The Unstored group is the control group, that is, 10% fetal bovine serum (FBS) is added to α-MEM to prepare a preservation solution, and then the freshly digested cells are suspended in the preservation solution to a cell density of 1×106 cells / ml, without preservation operation, directly to the experiment.

[0032] The compound electrolyte injection (ME) + human serum albumin (HSA) group is the mesenchymal stem cell injection of the present invention, that is, 5% HSA is added to the co...

Embodiment 2

[0053] Example 2: The mesenchymal stem cell injection prepared from a mesenchymal stem cell cryopreservation solution of the present invention was tested for proliferation ability, cell osteogenic differentiation ability test, cell adipogenic differentiation ability test and cell surface marker detection.

[0054] 2.1 Preparation of mesenchymal stem cell injection of the present invention

[0055] Add 5% HSA to the compound electrolyte injection to prepare a preservation solution, and then suspend freshly digested cells in the preservation solution to a cell density of 5×10 6 per ml, stored for 24 hours before experimenting.

[0056] The Unstored group is the above-mentioned Unstored group, which is the fresh cells digested after the cell preparation, and does not undergo cryopreservation. The Unstored group is the control group, which symbolizes the best state of the cells. This group is set to test whether the state of the cells changes significantly after our cryopreserva...

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Abstract

The invention discloses mesenchymal stem cell low-temperature preserving fluid and a preparation method thereof. The mesenchymal stem cell low-temperature preserving fluid is prepared from human albumin injection and compound electrolyte injection according to a volume ratio of (1:100) to (1:5). Per 25ml of human albumin injection contains 5g of human albumin; per 1000 mL of the compound electrolyte injection contains 5.26g of sodium chloride, 5.02g of sodium gluconate, 3.68g of sodium acetate, 0.37g of potassium chloride and 0.30g of magnesium chloride. The invention also discloses an injection prepared from the low-temperature preserving fluid and a preparation method thereof. The use of freeze storage protection agents having the toxic and side effects and causing injury on cells is avoided; the freeze storage and unfreezing steps are avoided; the clinic safety, simplicity and convenience are greatly improved; meanwhile, the activity maintenance time of stem cells is prolonged; the condition that the cell activity can still meet the clinic use requirements after the 24h transportation. The mesenchymal stem cell low-temperature preserving fluid has the advantages that the safety is high; the transportation is convenient; the cell activity is high; the cost is low.

Description

technical field [0001] The invention belongs to the technical field of stem cell preservation, and in particular relates to a mesenchymal stem cell cryogenic preservation solution and a preparation method thereof. The invention also relates to a mesenchymal stem cell injection prepared from the mesenchymal stem cell cryogenic preservation solution and a preparation method thereof . Background technique [0002] Mesenchymal stem cells [mesenchymal stem cells, MSC] are a type of stem cells derived from mesoderm with multi-directional differentiation potential, widely distributed in various tissues of the human body, such as bone marrow, fat, umbilical cord, placenta, amniotic membrane, amniotic fluid, menstrual blood, Dental pulp and pancreas etc. Mesenchymal stem cells can develop into bone, cartilage, fat, and other types of cells. Mesenchymal stem cells can support hematopoiesis and immune regulation, and have long-term development prospects in cardiovascular diseases, li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 张凤丽陈彦田赵鑫齐念民
Owner 浙江新生泉细胞科技有限公司
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