Microcarrier culture system of umbilical cord mesenchymal stem cells

A microcarrier and microcarrier technology, applied in tissue cell/virus culture devices, animal cells, tissue culture, etc., can solve the problems of MSC cell damage, high shear force, and difficulty in MSC collection.

Inactive Publication Date: 2013-09-18
侯亚义 +1
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Problems solved by technology

At present, there is no microcarrier culture system and its special medium suitable for large-scale culture of MSC, and the culture system for cultivating other cells and the general medium containing animal-derived serum are mostly used, which leads to the following problems: ①There are many apoptotic cells: this is mainly due to the large shear force generated by the commonly used microcarrier system under the stirring of an ordinary rotar

Method used

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  • Microcarrier culture system of umbilical cord mesenchymal stem cells
  • Microcarrier culture system of umbilical cord mesenchymal stem cells
  • Microcarrier culture system of umbilical cord mesenchymal stem cells

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example 3

[0029] Example 3 The application of MSC preservation solution for treatment:

[0030] 1. Preparation of three kinds of preservation solutions: in strict accordance with aseptic operation, take 250ml of clinical Ringer's solution, add 2.5ml of clinical human albumin and 250,000 AxaIU / ml of clinical heparin sodium, and divide into three parts evenly after mixing. Servings, 80ml each. Add separately to each serving: 10 -7 M estrogen and 50ng / ml of IL-1β (for the treatment of systemic lupus erythematosus), 50ng / ml of human Midkine (for the treatment of cardiovascular and cerebrovascular diseases) and 100ng / ml of human bone morphogenic protein BMP (for the treatment of osteogenesis).

[0031] 2. Add MSC cells to each preservation solution, the cell concentration is 5×10 6 cells / ml. After storage at 4°C for 24 hours, cell apoptosis was detected by flow cytometry.

[0032] 3. Use Q-PCR method to detect the expression of Cox-2, IL-8, IL-6 and CXCR4 factors after 24 hours of storag...

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Abstract

The invention belongs to the field of biological medicines, and discloses a microcarrier culture system of umbilical cord mesenchymal stem cells (MSC), and preparation and applications of an MSC preserving fluid. The microcarrier culture system is characterized by preparing a heparin crosslinking sephadex microcarrier and a mating microcarrier culture medium, and a high-efficiency MSC culture system can be built. MSC cells can be efficiently and fast prepared within a short period by utilizing the microcarrier culture system of the MSC. The MSC preserving fluid is characterized by adding human albumin, heparin sodium, estrogen and IL-1beta into a ringer solution, the cell activity of the MSC cells can be kept unchanged within 24 hours by utilizing the preserving fluid, and the level of cell factors can be improved by about five times on average. The microcarrier culture system is applied to being developed into large-batch preparation of umbilical cord MSC for clinical use and laboratories, and provides a specific preserving method for MSC for treating systemic lupus erythematosus, and establishes a basis for the clinical application of the MSC.

Description

technical field [0001] The present invention is a rapid, effective and large-scale method for obtaining umbilical cord MSCs, specifically, a serum-free and large-scale method for obtaining umbilical cord mesenchymal stem cells using cell microcarrier culture technology and special cytokines as a culture system. Yield, speed and quality, and develop special cell preservation solutions according to different clinical applications. technical background [0002] Mesenchymal stem cells (MSCs) are a type of pluripotent stem cells belonging to the mesoderm, which mainly exist in connective tissue and organ mesenchyme. Ease of acquisition and other characteristics have been widely valued clinically. In recent years, MSC has become a hot spot in stem cell research, and has good application prospects in the fields of immune disease treatment, hematopoietic stem cell transplantation, tissue engineering, and genetic engineering. However, the clinical application of MSC is limited by i...

Claims

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Application Information

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IPC IPC(8): C12N5/02C12N5/0775C12M3/02A01N1/02
Inventor 侯亚义赵树立曹志刚米琼宇樊竤冶
Owner 侯亚义
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