Activation method for stem cells based on colony stimulating factor 1

A colony-stimulating factor and activation method technology, applied in the direction of cell culture active agent, non-embryo pluripotent stem cells, biochemical equipment and methods, etc., can solve the problems of stem cell proliferation, slow differentiation of activated adult cells, etc., to promote mobilization, promote The effect of in vitro proliferation

Inactive Publication Date: 2016-06-29
佰通生物技术(苏州)有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of the above-mentioned deficiencies in the prior art, the purpose of the present invention is to provide a method for activating stem cells based on colony-stimulating factor 1, aiming to solve the problem that the prior art cannot solve the problem of slow proliferation, activation and differentiation of stem cells into other adult cells. The problem

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  • Activation method for stem cells based on colony stimulating factor 1
  • Activation method for stem cells based on colony stimulating factor 1
  • Activation method for stem cells based on colony stimulating factor 1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The mobilization effect of CSF1 on hematopoietic stem cells

[0037] 5-month-old mice were collected and subcutaneously injected with 1 microgram of CSF1 and 1 microgram of G-CSF per gram of body weight. The positive control was subcutaneously injected with 1 microgram of G-CSF per gram of body weight, and the negative control was injected with an equal volume of normal saline. Three days later, blood was collected from the tail vein in an anticoagulant tube added with heparin anticoagulant, and the number of hematopoietic stem cells in the peripheral blood after mobilization was traced by CD34 antibody conjugated with FITC, and CD34 positive cells were detected by flow cytometry quantity. The specific operation is as follows: 1) Wash the cells 3 times with PBS; 2) Dilute the antibody with PBS according to the antibody instruction manual, avoiding light; 3) Add the diluted antibody to the cells, and gently pipette to evenly form a single cell suspension , and incubate ...

Embodiment 2

[0039] Effect of CSF1 on Proliferation of Hematopoietic Stem Cells

[0040] Hematopoietic stem cells obtained by CD34 magnetic bead sorting were cultured in hematopoietic stem cell serum-free medium, and hematopoietic stem cells were inoculated into the culture bag and placed at 37°C (5% CO 2 ) culture, the culture bag was gently shaken every 12 hours, and the culture medium was supplemented with 2ng / mL CSF1 throughout the culture process. The cell density was visually measured under the microscope every 12 hours, and the total cell density was measured by MTT method after 72 hours. The results of the study show that under the medium conditions described in the present invention added with CSF1, the density of hematopoietic stem cells is 3.6 times higher than that obtained under the culture conditions without adding CSF1, and the data is the average value SEM of 5 independent experiments (P figure 2 shown).

Embodiment 3

[0042] The promotion effect of CSF1 on the proliferation of mesenchymal stem cells

[0043] The conventional medium formula for culturing mesenchymal stem cells is: DMEM / F12 basic medium, 10% fetal bovine serum (FBS). In order to compare the culture medium described in this example and to examine the influence of the culture medium described in this example on the expansion speed of human stem cell culture, this example selects human stem cells from three kinds of tissues and organs: fat, bone marrow and umbilical cord. Stem cells (ADSC, BMSC, UMBSC) are used for in vitro cell culture experiments, and the cell seeding density is controlled between 10-20%. Before seeding, the culture dish is treated with cell-adhesive materials at room temperature for 2 hours or incubated overnight at 4°C, and the culture medium is added after seeding Culture (37°C, 5% CO 2 ). Cell density was measured every 12 hours by MTT assay. The research results show that the expansion speed of hematop...

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Abstract

The invention discloses an activation method for stem cells based on a colony stimulating factor 1. The CSF1 is used for mobilizing, proliferating and activating the stem cells, and the CSF1 can promote in-vitro proliferation of the stem cells and promote mobilization, proliferation and activation of the stem cells in the body, and is expected to play an important role in cell culture, disease treatment and anti-aging healthcare.

Description

technical field [0001] The invention relates to the field of activation of stem cells, in particular to a method for activating stem cells based on colony-stimulating factor 1. Background technique [0002] A living body develops from a fertilized egg, after the formation of embryonic stem cells to tissues and organs and the acquired maintenance of mature individuals, after the formation of mature individuals, the number of stem cells in the body is very small, only about one ten thousandth of the cells in adult tissues It is stem cells, and it is these very small amount of stem cells that play a vital role in the acquired maintenance of life. Except for nerve cells that are immortal for life, the life span of most adult cells is only 100-300 days, but living organisms need to last for several years or even over 100 days. A hundred-year life span mainly depends on the proliferation, activation, and potential of stem cells to differentiate into other adult cells. [0003] Un...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074C12N5/0789C12N5/0775C12N5/077A61K49/00
CPCC12N5/0647A61K49/0008C12N5/0653C12N5/0654C12N5/0655C12N5/0663C12N5/0665C12N5/0667C12N2501/22
Inventor 刘小青
Owner 佰通生物技术(苏州)有限公司
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