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Cell sheet protection liquid

A technology of cell membrane and protective solution, which is applied in the direction of animal cells, vertebrate cells, cell culture active agents, etc., and can solve the problem of cell membrane integrity, cell connection damage in the membrane, affecting the clinical application of cell membranes, activity and secreted factors. issues such as guarantees

Active Publication Date: 2020-12-25
BOE TECH GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During this process, if the cell membrane preservation solution is not suitable, the activity and secretion factors of the cells in the cell membrane cannot be guaranteed, and the integrity of the cell membrane and the connection of cells in the membrane will also be destroyed, seriously affecting the integrity of the cell membrane. Clinical application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Example 1. Preparation of Mesenchymal Stem Cell Sheets

[0091] Before the preparation of the umbilical cord mesenchymal stem cell membrane, a layer of matrix that is conducive to the attachment of the mesenchymal stem cells is first coated on the surface of the temperature-sensitive culture dish. The matrix used was fibronectin at a concentration of 10 mg / L. The thermosensitive dishes were coated for 8 hours. Subsequently, the coating solution was discarded, and the single-cell suspension of the mesenchymal stem cells was added to the smart culture dish. After the cells proliferated for 12 hours after attachment, the temperature was lowered to 25°C. The cells will be detached from the bottom of the temperature-sensitive smart culture dish in sheets and layers, and become cell membranes that are completely connected with extracellular matrix. Among them, the diaphragm is off-white, with dense structure and smooth surface.

Embodiment 2

[0092] Example 2. Preparation of cell membrane protection solution

[0093] Exemplary cell sheet protection solutions with the following formulations A, B and C were prepared for further experiments.

[0094] Formulation A:

[0095] Buffered saline solution: PBS

[0096] Extracellular matrix component: gelatin (1mg / L)

[0097] Reducing agent: vitamin C (500mg / L); glutathione (10mg / L)

[0098] Substances needed for cell metabolism: transferrin (10mg / L); linoleic acid (1.0mg / L); linolenic acid (1.0mg / L)

[0099] Hormone substances: insulin (10mg / L)

[0100] Formulation B:

[0101] Buffered saline solution: Hanks buffer

[0102] Extracellular matrix component: fibronectin (10mg / L)

[0103] Reducing agent: vitamin C (500mg / L); sodium selenate (0.02mg / L)

[0104] Substances required for cell metabolism: transferrin (10mg / L); putrescine (10mg / L); human serum albumin (2000mg / L); carnitine (2mg / L)

[0105] Hormone substances: insulin (10mg / L); progesterone (0.01mg / L)

[01...

Embodiment 3

[0112] Example 3. Detection of cell secreted factors before and after transportation

[0113] The mesenchymal stem cell sheet prepared as described in Example 1 was placed in the cell sheet protection solution with formula A, formula B and formula C, and transported at 4° C. for 48 hours. Exemplary photos of cell membranes before and after transportation in the cell membrane protection solution of formula B are as follows figure 1 shown. It can be seen from the photo that before and after transportation, the cell membrane has maintained its integrity, its shape has not changed significantly, and it has the characteristics of smooth edges, flat surface, and no looseness or wrinkles.

[0114] The therapeutic effect of mesenchymal stem cells on self-tissue repair and immune-related diseases is mainly achieved by secreting and expressing immunosuppressive factors, cytokines, growth factors, etc. to regulate inflammation and tissue homeostasis. In this example, the cytokine HGF...

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Abstract

The invention relates to the field of tissue engineering and regenerative medicine. Specifically, the invention provides a cell sheet protection liquid capable of maintaining the activity and cytokinesecretion ability of cells in a cell sheet during storage and transportation of the cell sheet, maintaining the integrity of the cell sheet and cell junction in the sheet, and protecting the cells from apoptosis. The cell sheet protection liquid comprises the following components: a, a buffer salt solution, b, an extracellular matrix component, c, a reducing agent, d, a substance required by cellmetabolism, and e, a hormonic substance. The invention also relates to the use of the cell sheet protection liquid in the storage and transportation of cell sheets.

Description

technical field [0001] The present disclosure relates to the field of tissue engineering and regenerative medicine, in particular to a cell membrane protection solution and its use in storing and transporting cell membranes. [0002] technical background [0003] At present, the incidence of tissue and organ damage caused by degenerative diseases or trauma (such as heart failure, Parkinson's disease, Alzheimer's disease, etc.) is increasing year by year, and traditional medical methods are difficult to completely cure these diseases. Stem cells include pluripotent stem cells and adult stem cells, which have the potential to differentiate into a variety of functional cells, and can also secrete a variety of cytokines that are beneficial to the repair of tissues and organs. They are ideal cells for repairing damaged tissues and organs. [0004] At present, there have been a large number of basic research and clinical research on stem cells repairing damaged tissues and organs i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0226C12N5/0668C12N2533/52C12N2533/54C12N2523/00C12N2500/38C12N2500/32C12N2500/33C12N2500/25C12N2501/392C12N2500/30A61L27/3834C12N5/0662C12N2539/10
Inventor 王娟赵玉菲高爽靳新谭玉琴李志生常德华
Owner BOE TECH GRP CO LTD
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