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Method for treating sclerosis using placental mesenchymal stem cell preparation

A technology of cell preparation and mesenchymal stem cells, applied in the field of biotechnology and biomedicine

Active Publication Date: 2019-04-19
BOYALIFE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods are subject to further improvement in terms of extract purity and / or recovery

Method used

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  • Method for treating sclerosis using placental mesenchymal stem cell preparation
  • Method for treating sclerosis using placental mesenchymal stem cell preparation
  • Method for treating sclerosis using placental mesenchymal stem cell preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0192] Embodiment 1, placental whole cell processing:

[0193] 1. Pre-preparation of the mixed synthase digestion solution: pipette 22ml of HBSS (Hank's Balanced Salt Solution) containing calcium and magnesium ions, 0.4ml of Roche Liberase MNP-S enzyme (for example, purchased from Xibao Biology, article number: 5578582001), 0.7ml Put ml of DNA type I enzyme in a 50ml centrifuge tube, then add zinc chloride (addition concentration is 0.2g / L, 0.25g / L, or 0.3g / L), mix well, and preheat at 37°C for more than 20min. The Hank's balanced salt solution consists of: 8.0g / L of NaCl, 0.4g / L of KCl, 0.1g / L of MgSO4 7H2O, 0.1g / L of MgCl2 6H2O, 0.06g / L of Na2HPO4 2H2O, 0.06g / L of KH2PO4, 1.0g / L of glucose, 0.14g / L of CaCl2, 0.35g / L of NaHCO3, 0.2g / L of phenol red, hydrochloric acid or sodium hydroxide to adjust the pH to 7.4.

[0194] 2. Preparation of placenta lobule: take out the placenta from the collection bag and place it in a white porcelain plate, rinse with tissue cleaning soluti...

Embodiment 2

[0211] Embodiment 2, primary cell recovery and subculture

[0212] 1. Cell recovery:

[0213] Take 2 tubes of frozen cells, thaw them quickly at 37°C, transfer the cells to a 15ml centrifuge tube, add 8ml of complete medium to recover by dripping; unless otherwise stated, the complete medium used in this article is DMEM-F12 culture containing 10% FBS base;

[0214] Then centrifuge at 1200rpm for 5min (acceleration 9, deceleration 7), remove the supernatant, add 5ml of complete medium to resuspend; each tube of cells is inoculated in a T75 culture flask, supplement the complete medium to 30ml, and place in a CO2 incubator ( Culture at 37°C, 5% CO2, saturated humidity); replace the medium with complete medium every 3-4 days, count according to the colony formation after 12 days of recovery, until the cell density is not less than 3000 cells / cm 2 Subsequent passages can be carried out;

[0215] 2. Cell subculture: Take the revived P0 generation cells and wash them with PBS, ...

Embodiment 3

[0229] Embodiment 3, biological characteristic identification of placental MSC

[0230] With reference to the methods [0062] to [0089] of the authorized patent CN102676451A, the biological characteristics of placental mesenchymal stem cells were identified. The ability of cell differentiation proves that the MSC obtained by the method of the present invention has stem cell characteristics.

[0231] For example, exemplarily, the induction differentiation test is performed on the P5 generation cells, and the results show that these cells have the ability to differentiate into osteoblasts, adipocytes, and chondrocytes. Typical micrographs of adipogenic, osteogenic, and chondrogenic differentiation are shown in Figure 6 .

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Abstract

The invention relates to a method for treating scleroses using a placental mesenchymal stem cell preparation. Particularly, on one hand, the invention relates to the use of the cell preparation for the manufacture of a medicament for the treatment and / or prevention of systemic sclerosis; the cell preparation is a cell suspension prepared by suspending mesenchymal stem cells such as placental mesenchymal stem cells in a 0.9% sodium chloride solution, and the cell preparation has a cell concentration of 1-10*10<6> cells / ml. On the other hand, the invention also relates to a cell preparation forthe treatment and / or prevention of systemic sclerosis, and a method for preparing the cell preparation, the method for preparing mesenchymal stem cells and the mesenchymal stem cells are involved in the cell preparation, the mixed enzyme digest is used in these methods. The cell preparation has excellent biological effect in the aspect of treating scleroderma.

Description

technical field [0001] The invention belongs to the fields of biotechnology and biomedicine, and relates to a method for treating sclerosis with a placental mesenchymal stem cell preparation and the placental mesenchymal stem cell preparation used in the method. Specifically, the present invention relates to a method for isolating stem cells from the placenta, in particular to a method for isolating mesenchymal stem cells from the placenta, and more particularly relates to a digestive enzyme composition using the unique formula of the present invention to extract from the placenta A method for isolating mesenchymal stem cells from tissues and culturing them into mesenchymal stem cells. Using the method of the invention can effectively improve the efficiency of isolating mesenchymal stem cells from placenta. Further, the present invention relates to using the above-mentioned placental mesenchymal stem cells to prepare cell preparations, and then use the cell preparations to tr...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61P37/00A61P17/00C12N5/0775
CPCA61K35/28A61P17/00A61P37/00C12N5/0668C12N2509/00
Inventor 许晓椿李容肖海蓉刘冰
Owner BOYALIFE
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