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Method for storing mesenchymal stem cells and method for culturing mesenchymal stem cells

A technology of mesenchymal stem cells and preservation methods, applied in the field of preservation methods and cultivation of mesenchymal stem cells, to achieve the effect of reducing operating costs

Active Publication Date: 2011-06-29
SHANDONG QILU STEM CELL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for preserving umbilical cord mesenchymal stem cells, so as to overcome the limitation that expansion must be carried out before storing umbilical cord mesenchymal stem cells in the prior art

Method used

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  • Method for storing mesenchymal stem cells and method for culturing mesenchymal stem cells
  • Method for storing mesenchymal stem cells and method for culturing mesenchymal stem cells
  • Method for storing mesenchymal stem cells and method for culturing mesenchymal stem cells

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Embodiment 1

[0033] The umbilical cord used was the product of normal labor, and the mother's consent was obtained. No intrauterine hypoxia occurred during delivery, the color of the umbilical cord was normal ivory white, and there was no hyperemia. The umbilical cords tested negative for hepatitis A, hepatitis B, syphilis and AIDS can be separated, frozen and cultured in the next step.

[0034] Prepare cryopreservation solution (CL solution). Firstly, prepare cryopreservation and resuscitation base solution, which is low-sugar DMEM (Dulbecco's modified Eagle's medium) medium (LG-DMEM) containing 50% (volume / volume; mL / mL) fetal bovine serum. Then add 18.6% (volume / volume; mL / mL) of DMSO, 15.4% (mass / volume; g / mL) of acetamide, 9.0% (volume / volume; mL / mL) of 1,2-propanediol and 4.0% (mass / volume; g / mL) polyethylene glycol (molecular weight: 6000) were used to make CL solution.

[0035] All liquids were sterilized using 0.22 μm pore size filters.

[0036] The collection and transportati...

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Abstract

The invention provides a method for storing mesenchymal stem cells and a method for culturing mesenchymal stem cells. The storing method comprises the following steps of: cutting the umbilical cord into blocks, and putting the blocks into a mixed solution of freezing storage solution and base solution in a volume rate of 1:3 for pretreatment; putting the blocks in the mixed solution of the freezing storage solution and the base solution in a volume rate of 1:1 for treatment at the temperature of 0 DEG C; putting the blocks in the freezing storage solution for treatment at the temperature of 0DEG C; and freeze-storing the blocks in liquid nitrogen. The culturing method is that the stored umbilical cord blocks are used for culture. After babies are delivered, the expensive cell culturing operation of the umbilical cords is not required immediately, the umbilical cords can be directly frozen at the deep low temperature, and the cell activity is not affected. One freezing storage tube can be picked up at any time, the umbilical cords are recovered and successfully cultured and amplified to obtain MSCs, so that the culture operation is not required to be made for each portion of umbilical cord, and the operating cost of the umbilical cord MSCs storage warehouse is greatly lowered.

Description

technical field [0001] The invention belongs to the technical fields of cell biology and low temperature medicine, and in particular relates to a preservation method and a culture method of mesenchymal stem cells. Background technique [0002] Mesenchymal Stem Cells (MSCs) have broad application prospects in developmental biology research and clinical treatment, such as tissue and organ repair (Blood, 102, 3483-3493, 2003). MSCs were first confirmed in the bone marrow, and then MSCs were found in adult peripheral blood, periosteum, muscle and adipose tissue. MSCs belong to pluripotent stem cells, which can differentiate into different types of cells such as bone, fat, cartilage, muscle and liver cells under corresponding induction conditions (Science, 284, 143-147, 1999). The umbilical cord is a rich source of normal childbirth waste, which is rich in MSCs. Recently, there have been many reports on the isolation of MSCs from umbilical cords (Stem Cells, 22, 1330-1337, 2004...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N5/0775
Inventor 李栋侯怀水肖琼范奉群刘东
Owner SHANDONG QILU STEM CELL ENG
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