Purpose of umbilical cord mesenchymal stem cell exosome

A technology of mesenchymal stem cells and exosomes, applied in the field of stem cell culture, can solve problems such as unsatisfactory hematopoietic stem cell effects and hematopoietic stem cell damage

Inactive Publication Date: 2017-02-15
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cell mobilization will cause damage to hematopoietic stem cells, which has great side effects
[0004] If a large number of peripheral hematopoietic stem cells can be multiplied in vitro, it will provide more options for stem cell transplantation, but at present, the effect of in vitro expansion of hematopoietic stem cells is not ideal

Method used

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  • Purpose of umbilical cord mesenchymal stem cell exosome
  • Purpose of umbilical cord mesenchymal stem cell exosome
  • Purpose of umbilical cord mesenchymal stem cell exosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0042] Example 2 Isolation and purification of human peripheral blood hematopoietic stem cells:

[0043] About 5 mL of peripheral blood was used to separate bone marrow mononuclear cells by density gradient centrifugation. The specific method is as follows: add the collected bone marrow samples to an equal amount of PBS, add slowly, centrifuge at 4°C, and centrifuge at 2000 rpm for 30 minutes. The milky white intermediate layer in the middle is drawn, which is the layer rich in monocytes. The obtained monocytes were resuspended in IMDM complete medium (IMDM+10%FBS+1%PS+6ng / ml IL-3+10ng / ml SCF+10ng / ml IL-6), pipetted evenly, and the cells were count. cells in 1x10 6 The density of / mL was inoculated in a 100mm Petri dish and placed at 37°C with a volume fraction of 5% CO. 2 Incubate for 24h.

[0044] Separation of CD34 from cultured peripheral blood mononuclear cells by magnetic bead sorting + Hematopoietic stem cells (HSCs). The specific method is as follows: the isolat...

Embodiment 3

[0046] test group:

[0047] The HSCs prepared in Example 2 were 1×10 6 Cell / mL density was seeded in 24-well plate with 1mL of medium (IMDM+10%FBS+1%PS+6ng / ml IL-3+10ng / ml SCF+10ng / ml IL-6), and 0.5mL of umbilical cord MSCs was added Exosomes, the volume ratio of medium to exosomes is 2:1, placed in 5% CO 2 The cells were cultured at 37°C in an incubator for 3 days. After 3 days, they were mixed with 0.4% trypan blue at a volume ratio of 1:1, observed and counted under a microscope, and cell viability was calculated according to the staining situation.

[0048] Cells were cultured at 6 × 10 3 The density of inoculated in 0.3mL IMDM complete medium, transferred to Methocult TM In H4531 medium, after mixing, let stand for 5 minutes at room temperature. Transfer 1.1mL of cell and medium suspension to a 35mm cell culture dish and place in 5% CO 2 The cells were cultured at 37°C in an incubator, and the colony formation ability was observed after 7 days, and the cells were ph...

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Abstract

The invention relates to the field of stem cell culture, and particularly relates to a purpose of umbilical cord mesenchymal stem cell exosome. Shown by experiments, with respect to culture experiments adopting a complete medium, by adopting co-culture of the umbilical cord mesenchymal stem cell exosome and peripheral blood stem cells, the amount of cells acquired by culture can be increased by about 1.5 times, and a clone formation amount is also increased by about 1.5 times. Through statistical analysis, compared with a culture effect without the umbilical cord mesenchymal stem cell exosome, the experiment effect with addition of the umbilical cord mesenchymal stem cell exosome has an extremely remarkable difference (p is smaller than 0.001).

Description

technical field [0001] The invention relates to the field of stem cell culture, in particular to the use of umbilical cord mesenchymal stem cell exosomes. Background technique [0002] Peripheral hematopoietic stem cells are hematopoietic stem cells present in peripheral blood. In 1979, Goldman et al. collected cryopreserved peripheral blood cells for a group of chronic myeloid leukemia patients in the accelerated phase or blast transition phase at the initial diagnosis, so that the patients returned to the chronic phase, and began the clinical application of peripheral blood hematopoietic stem cell transplantation (PBSCT). [0003] Under normal circumstances, the content of stem / progenitor cells in peripheral blood accounts for 0.01% to 0.1% of mononuclear cells, which is equivalent to 1% to 10% of bone marrow. After mobilization, CD34 in peripheral blood MNCs + The number of cells and CFU were significantly higher than those of bone marrow. So far, it has been found that...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789C12N5/0775
CPCC12N5/0647C12N5/0668
Inventor 陈海佳葛啸虎王一飞冯潜张维敏
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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