A method for promoting the proliferation of human bone marrow mesenchymal stem cells based on exosomes

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

Active Publication Date: 2020-07-17
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 amniotic mesenchymal stem cells on promoting the growth of HDF cells. Simple analogy to other sources of exosomes to promote the role of other types of cells
Therefore, whether the exosomes secreted by stem cells can be used to promote the proliferation and growth of BMSCs in vitro is currently not publicly reported in any literature or patents.

Method used

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  • A method for promoting the proliferation of human bone marrow mesenchymal stem cells based on exosomes
  • A method for promoting the proliferation of human bone marrow mesenchymal stem cells based on exosomes
  • A method for promoting the proliferation of human bone marrow mesenchymal stem cells based on exosomes

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Experimental program
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Effect test

Embodiment 1

[0038] Example 1: Separation 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 medium (product of Dako), trypsin (product of Gibco), BCA protein quantification kit (Beyond), 5XSDS loading buffer, CD63 Rabbit anti-human antibody (BD pharmingen), CD9 rabbit anti-human antibody (BD pharmingen), HRP-labeled goat anti-rabbit IgG secondary antibody (Tiangen company), ECL chemiluminescent detection kit (Beiyuntian company).

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

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

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

[0044] After harvestin...

Embodiment 2

[0052] Embodiment 2: cell proliferation experiment

[0053] 1. Cell count

[0054] Human bone marrow mesenchymal stem cells (bone marrow primary culture, bone marrow from the Department of Orthopedics, Changzheng Hospital, preparation method reference: Majumdar, M.K., Banks, V., Peluso, D.P., and Morris, E.A. (2000) Isolation, characterization, and chondrogenic potential of human bone marrow-derived multipotential stromal cells. Journal of cellular physiology. 185, 98-106) were inoculated into 96-well plates at a concentration of 1000 cells per well, and then added human umbilical cord mesenchymal stem cells after quantitative protein concentration Secretosomes were added according to the final concentration of 10 μg / ml and 20 μg / ml in the culture medium respectively, which were set as 10 μg / ml group and 20 μg / ml group, and a blank control group was set at the same time, set at 6h, 12h, and 24h , 36h four counting time points, set three wells for each time point in each group...

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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 in promoting 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] Mesenchymal stem cells (MSCs) are a type of pluripotent stem cells with self-renewal and multi-directional differentiation potential, which can differentiate into various tissues and cells after induction. It is an ideal seed cell in regenerative medicine (see literature: Dominici, M., Le Blanc, K., Mueller, I., Slaper-Cortenbach, I., Marini, F., Krause, D., Deans, R., Keating, A., Prockop, D., and Horwitz, E. (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 8, 315-317). Human bone marrow mesenchymal st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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