Umbilical cord mesenchymal stem cells for treating lung diseases and preparation method thereof

A technology of stem cells and umbilical cord, applied in the field of biomedicine, can solve the problems of cumbersome operation and complex components

Pending Publication Date: 2020-08-11
SHENZHEN HORNETCORN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method is cumbersome to operate and h...

Method used

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  • Umbilical cord mesenchymal stem cells for treating lung diseases and preparation method thereof
  • Umbilical cord mesenchymal stem cells for treating lung diseases and preparation method thereof
  • Umbilical cord mesenchymal stem cells for treating lung diseases and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Embodiment 1 A kind of preparation method of umbilical cord mesenchymal stem cells

[0066] The umbilical cords were obtained from healthy full-term pregnancy cesarean section fetuses, and were isolated and cultured within 12 hours after collection using the tissue block adherence method. The specific steps are as follows:

[0067] (1) Cut the blood vessel longitudinally along the lumen of the umbilical vein, peel off the umbilical vein intima, and cut the remaining tissue into 3-5mm 3 Small pieces, transferred to a T75 cell culture flask, added 10mL of complete medium (DMEM / F12 medium containing 2% serum substitute) to make it evenly distributed, placed at 37°C, 5% CO 2 , saturated humidity CO 2 in the incubator.

[0068] (2) Change the medium in full amount on the 7th day, and change the medium in half on the 12th day. Count the number of colonies on the 14th day of culture. More than 50 cells are counted as colonies. When the cells reached 70%-80% confluence, dige...

Embodiment 3

[0084] Embodiment 3 Lymphocyte suppression experiment result

[0085] In this example, the human umbilical cord mesenchymal stem cells cultured in Example 1 were used as the research object, the inhibitory effect of umbilical cord mesenchymal stem cells on peripheral blood mononuclear cells (PMBC) was detected by the CCK8 method, and the umbilical cord mesenchymal stem cells were evaluated by flow cytometry. The effect of mesenchymal stem cells on peripheral blood mononuclear cells, while analyzing its role in immune regulation. The results show that umbilical cord mesenchymal stem cells have an immunoregulatory effect on lymphocytes, can promote the production of Treg cells and inhibit the secretion of inflammatory factors such as IL-17.

[0086] (1) CCK8 detects the inhibitory effect of umbilical cord mesenchymal stem cells on peripheral blood mononuclear cells

[0087] The results after P2 generation umbilical cord mesenchymal stem cells inhibited peripheral blood mononucl...

Embodiment 4

[0095] Example 4 Detection of the ability of inflammatory factors to stimulate human umbilical cord mesenchymal stem cells to secrete indoleamine 2,3-dioxygenase

[0096] This example is carried out using the human umbilical cord mesenchymal stem cells obtained in Example 1.

[0097] Umbilical cord mesenchymal stem cells of P1 and P5 generations were used as research objects, and the expression of indoleamine 2,3-dioxygenase in umbilical cord mesenchymal stem cells stimulated by TNF-α and INF-γ was detected by ELISA, and its expression was evaluated. Immunomodulatory effect. The results show( Figure 4 ), P1 and P5 generation cells compared with the control group, the concentration of indoleamine 2,3-dioxygenase in the stimulation group was significantly increased, P<0.05; indicating that the cells can play a good immune regulation function in the inflammatory environment. It shows that the cells have good paracrine function, and the P1 and P5 generation cells have the same ...

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Abstract

The invention provides umbilical cord mesenchymal stem cells for treating lung diseases and a preparation method thereof. The umbilical cord mesenchymal stem cells provided by the invention can be used for preparing drugs for preventing or treating pneumoconiosis injury and fibrosis. The drugs are high in safety and good in intervention and treatment effect on corresponding diseases when being applied to lung disease treatment, especially pneumoconiosis lung injury and fibrosis thereof, quantitative production can be achieved through mature cell culture technical means, and the medicine has bright application prospects.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to an umbilical cord mesenchymal stem cell for treating lung diseases and a preparation method thereof. Background technique [0002] Stem cells have the characteristics of self-renewal ability and multi-directional differentiation potential. Mesenchymal stem cells (mesenchymal stem cells, mesenchymal stem cells) are one of the important members of the stem cell family. They originate in the early mesoderm and ectoderm, and first in the bone marrow. It was discovered, and then its characteristics of multidirectional differentiation, hematopoietic assistance, and immune regulation were gradually discovered. Whether in vivo or in vitro, mesenchymal stem cells can be induced and differentiated into various tissue cells. [0003] In 2001, Kotton proposed that adult tissue-derived mesenchymal stem cells could home to the lungs, which brought new opportunities for stem cells to trea...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61P11/00
CPCA61K35/28A61P11/00C12N5/0665C12N2509/00
Inventor 王宇环罗晓玲张正涵
Owner SHENZHEN HORNETCORN BIOTECH
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