Method for promoting human bone mesenchymal stem cell proliferation based on exosome

A bone marrow mesenchymal stem cell technology, applied in the field of promoting the growth of human bone marrow mesenchymal stem cells, can solve different problems and achieve stable properties

Active Publication Date: 2016-02-24
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in view of the fact that the components of exosomes secreted by stem cells from different sources and even different culture methods are significantly different, and the medium formulations and growth-dependent conditions of different cells are completely different, it is impossible to report the effect of exosomes from amnio...

Method used

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  • Method for promoting human bone mesenchymal stem cell proliferation based on exosome
  • Method for promoting human bone mesenchymal stem cell proliferation based on exosome
  • Method for promoting human bone mesenchymal stem cell proliferation based on exosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Isolation and purification of umbilical cord mesenchymal stem cells

[0039] The main materials and sources used are as follows:

[0040] Reagents: low-sugar DMEM (product of gibco), mesenchymal stem cell culture medium (product of Dakko), trypsin (product of gibco), BCA protein quantification kit (biyuntian), 5XSDS loading buffer, CD63 Rabbit anti-human antibody (BDpharmingen company), CD9 rabbit anti-human antibody (BDpharmingen), HRP-labeled goat anti-rabbit IgG secondary antibody (Tiangen company), ECL chemiluminescence detection kit (Biyuntian company).

[0041] Instruments: inverted microscope (Olympus), carbon dioxide incubator (Thermo), clean bench, benchtop centrifuge, transmission electron microscope (Philips), chemiluminescence gel imaging system (bioshine).

[0042] The umbilical cord comes from the Obstetrics and Gynecology Department of Changhai Hospital.

[0043] 1. Cultivation of human umbilical cord mesenchymal stem cells:

[0044] After obtaining the ...

Embodiment 2

[0052] Example 2: Cell Proliferation Experiment

[0053] 1. Cell count

[0054] Human bone marrow mesenchymal stem cells (primary culture of bone marrow, bone marrow from the Orthopedics Department of Changzheng Hospital, preparation method reference: Majumdar, MK, Banks, V., Peluso, DP, and Morris, EA (2000) Isolation, characterization, and chondrogenic potential of human bonemarrow- derivedmultipotentialstromalcells.Journalofcellularphysiology.185,98-106) were seeded into 96-well plates at a concentration of 1000 cells per well, and then human umbilical cord mesenchymal stem cell exosomes with quantitative protein concentration were added, and the final quantification was as follows: Add the final concentration of 10μg / ml and 20μg / ml, set them to the 10μg / ml group and 20μg / ml group, and set the blank control group at the same time, set 6h, 12h, 24h, 36h four counting time points, each group Set three wells at the time point and place the cells at 37°C, 5% CO 2 Cultivate in a car...

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Abstract

The invention relates to the field of biotechnology and mainly relates to a use of an early-generation human umbilical cord mesenchymal stem cell exosome in promotion of human bone mesenchymal stem cell growth. The use method is a method for promoting bone mesenchymal stem cell growth. The invention also relates to an extraction method of a human umbilical cord mesenchymal stem cell exosome. The early-generation human umbilical cord mesenchymal stem cell exosome and human bone mesenchymal stem cells are co-cultured so that human bone mesenchymal stem cell growth is obviously promoted, cell doubling time is shortened and a ratio of an S period in a cell period is increased.

Description

Technical field [0001] The invention relates to the field of biotechnology, and mainly relates to the application of early-generation human umbilical cord mesenchymal stem cell exosomes to promote the growth of human bone marrow mesenchymal stem cells, that is, a method for promoting the growth of human bone marrow mesenchymal stem cells. Background technique [0002] Mesenchymalstemcells (MSCs) are a type of pluripotent stem cells with self-renewal and multi-differentiation potential. After induction, they can differentiate into a variety of tissues and cells. Because of their convenient materials and no immune rejection, they are regeneration An ideal seed cell in medicine (see literature: Dominici, M., LeBlanc, K., Mueller, I., Slapper-Cortenbach, I., Marini, F., Krause, D., Deans, R., Keating, A., Prockop, D., and Horwitz, E. (2006) Minimal criteriaford efining multipotent mesenchymalstromal cells. The International Society for Cellular Therapy position statement. Cytotherapy...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28
Inventor 严冰浩方硕许震宇刘厚奇王越
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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