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Decidua parietalis mesenchymal stem cell culture medium for treating tumors and co-culture method

A technique for MSCs and tumor cells, applied in the field of co-cultivation, parietal decidua mesenchymal stem cell culture medium, which can solve the direct analysis of rare MSC homing ability and tumor type selectivity and adaptability, and the ability of parietal decidua to directly analyze Insufficient research on mesenchymal stem cells and tumors, and different reports on the impact of malignant tumor cells on biological behavior

Active Publication Date: 2021-08-31
GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. In the research methods of MSCs and tumors in the relevant literature, the effect of MSCs on a single tumor is mostly seen, and the direct analysis of the homing ability of MSCs and the selectivity and adaptability of tumor types is rarely seen;
[0006] 2. In the research methods of MSCs and tumors in the relevant literature, it is common to study the combination of modified mesenchymal stem cells and drugs on tumors, and the research on maternal parietal decidual mesenchymal stem cells and tumors is insufficient;
[0007] 3. In the co-culture method in the relevant literature, the research concentration used by PMSCs is single, resulting in different reports on the impact on the biological behavior of malignant tumor cells

Method used

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  • Decidua parietalis mesenchymal stem cell culture medium for treating tumors and co-culture method
  • Decidua parietalis mesenchymal stem cell culture medium for treating tumors and co-culture method
  • Decidua parietalis mesenchymal stem cell culture medium for treating tumors and co-culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] A method for preparing parietal decidual mesenchymal stem cells, comprising the following steps:

[0062] S11. Separation of parietal decidua tissue: use tissue cleaning solution to clean the blood and blood clots on the placental tissue of healthy full-term newborns, use surgical stripping instruments to separate parietal decidua tissue, and then use surgical scissors to cut it into 1-4mm 3 and cleaned with tissue cleaning solution; the tissue cleaning solution is prepared from the following volume percentage raw materials: 1% penicillin and streptomycin mixture, 51.1% erythrocyte lysate, 47% normal saline, and the normal saline is the mass fraction 0.9%;

[0063] S12. Digestive wall decidua tissue block: After adding tissue digestive juice to the chopped tissue block, shake and digest at a constant temperature for 2 hours at 37°C, with a rotation speed of 200rpm / min; the tissue digestive juice contains a volume concentration of 50% Tryple- EDTA enzyme and 10mg / ml typ...

Embodiment 2

[0071] The recovery method of the parietal decidual mesenchymal stem cell culture medium, including:

[0072] S21. Dissolving the frozen decidual mesenchymal stem cells cryopreserved in Example 1 in a water bath at 37°C;

[0073] S22. Use the selection medium to resuspend the epithelial decidual mesenchymal stem cells of step S21, centrifuge at 1300rpm for 6min, wash the cell surface twice with PBS buffer, centrifuge at 1300rpm for 6min, add the selection medium to resuspend the pellet for subculture;

[0074] S23. Collect the PMSCs of the P5 generation, add selection medium to resuspend the cells into a low concentration group, a medium concentration group and a high concentration group, wherein the concentration of PMSCs in the low concentration group is 1×10 6 per well, the concentration of PMSCs in the middle concentration group was 2×10 6 per well, the concentration of PMSCs in the high concentration group was 4×10 6 cells / well, each group was plated in the upper chambe...

Embodiment 3

[0076] Co-cultivation experiment of parietal decidual mesenchymal stem cell medium and cervical cancer cells, including:

[0077] S31. Dissolve the cervical cancer cell line Hela rapidly in a 37°C water bath, add complete medium to resuspend the cells, centrifuge, wash twice with PBS buffer, centrifuge at 1000rpm for 3min, add fresh complete medium to resuspend the cells, transfer Cultivate in a T25 culture bottle, which is designated as the P1 generation; the complete medium is a serum-free DMEM medium containing 10% fetal bovine serum and 1% anti-penicillin and streptomycin;

[0078] S32, Hela cell passage: observe the cell morphology and medium changes after recovery, replace the complete medium every 2 days, and collect the cell suspension when the cell growth and confluence is greater than 80%, count and calculate the survival rate, microbiological detection, and passage ;

[0079] S33. Cultivate Hela cells in the lower chamber of the Transwell chamber: Discard the origi...

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Abstract

The invention discloses a decidua parietalis mesenchymal stem cell culture medium for treating tumors and a co-culture method. The decidua parietalis mesenchymal stem cell culture medium for treating tumors contains decidua parietalis mesenchymal stem cells and a selective culture medium, the decidua parietalis mesenchymal stem cells have a homing effect, a proliferation inhibiting effect and a cell cycle adjusting effect on various tumor cells.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a medium for treating tumors and a co-culture method for parietal decidual mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSC) are pluripotent progenitor cells with the ability to chemoattract tumors, and the research on tumors has made significant progress. However, there are different reports on the influence of uterine-derived mesenchymal stem cells on the biological behavior of gynecological malignant tumor cells. The study of the adaptability and selectivity of parietal decidual MSCs (Pariduval Mesenchymal stem cells, PMSCs) and tumors has become a new clinical treatment for tumors. method. [0003] Pariduval Mesenchymal stem cells (PMSCs) have the characteristics of simple in vitro proliferation and culture, no moral ethics involved, good migration ability, homing to tumor cells and low immunogenicity, and avoid bone marrow The difficulty in obt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/09A61P35/00A01N1/02
CPCC12N5/0668A61P35/00A01N1/0221C12N2501/115C12N2501/11C12N2501/125C12N2509/00C12N2509/10C12N2502/30
Inventor 刘小翠唐淑艳邓燕莲蒙燕瑶雅思敏杨景利王进辉
Owner GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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