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Hematopoietic stem cell culture medium and its application and stem cell cultivation method based on hematopoietic stem cell culture medium

A technology for hematopoietic stem cells and culture medium, applied in cell culture medium, cell culture active agents, biochemical equipment and methods, etc., can solve the problems of slow proliferation rate and low passage number of hematopoietic stem cells.

Inactive Publication Date: 2015-08-12
SHENZHEN FULIXIN HEALTH IND DEV LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a medium for hematopoietic stem cells and its application and a method for culturing stem cells, aiming to solve the problems of relatively slow proliferation of hematopoietic stem cells and low number of passages in the prior art

Method used

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  • Hematopoietic stem cell culture medium and its application and stem cell cultivation method based on hematopoietic stem cell culture medium
  • Hematopoietic stem cell culture medium and its application and stem cell cultivation method based on hematopoietic stem cell culture medium
  • Hematopoietic stem cell culture medium and its application and stem cell cultivation method based on hematopoietic stem cell culture medium

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preparation example Construction

[0127] S100. Preparation of culture medium: adding amino acids, vitamins, salts, lipids, protein polypeptides and cytokines to the culture medium in sequence. Of course, for step S100, the substances to be added depend on the actual situation. For example, plant or animal cell extracts can be added. The specific process is to undergo high-temperature extraction treatment, and high-speed centrifugation to remove large insoluble particles, and then Filter with 0.22 μm and 0.1 μm filter membranes successively, inactivate and remove insoluble precipitates and potential pathogens, and then add to the culture medium. In addition, trace elements or small organic molecules can also be added.

[0128] S200. Preparation of hematopoietic stem cells: removing anucleated or multinucleated cells from peripheral blood or umbilical cord blood, and isolating CD34+ cells. Specifically, a certain amount of serum or plasma is added to the culture flask, and anucleated or multinucleated cells are...

Embodiment 1

[0143] Example 1. Proliferation of hematopoietic stem cells

[0144] In order to check the influence of the hematopoietic stem cell culture medium of the present invention on the expansion speed of hematopoietic stem cells, the peripheral blood after 4 days of G-CSF mobilization was selected to prepare mononuclear cells with the Ficol method, and the obtained monocytes were subjected to in vitro cell culture experiments. cells by 10 4 Add control culture medium (control group) and the hematopoietic stem cell culture medium (experimental group) of the present invention after inoculation and cultivate respectively (37 degrees Celsius, 5% CO 2 ), 4-7 days later, hematopoietic stem cells characterized by CD34+ were sorted by anti-CD34 antibody flow cytometry, and the number of CD34+ cells and their ratio to the total cells were calculated, and the total number of cells was determined by the MTT method.

[0145] The results show that under the same conditions, the CD34+ cell ratio...

Embodiment 2

[0146] In order to check the influence of the hematopoietic stem cell culture medium of the present invention on the telomere length of hematopoietic stem cells, after injecting mice with G-CSF and mobilizing them for 4 days, get peripheral blood to prepare mononuclear cells by the Ficol method, and further use conjugated anti- Magnetic beads of CD34 antibody enrich CD34+ cells. Divide CD34+ cells by 10 4 Inoculated into the hematopoietic stem cell medium (experimental group) of the present invention and conventional control medium (control group), suspension culture (37 degrees Celsius, 5% CO 2 ) After 1 week, the telomere length of the cultured cells was measured according to the fluorescent quantitative PCR method described in the literature (Journal of Clinical and Experimental Medicine 2008(7):14). The results showed that the telomere length of the experimental group was longer than that of the control group 8% (see Figure 4 ); data are mean±SEM of 12 independent exper...

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Abstract

The invention discloses a hematopoietic stem cell culture medium and its application and a stem cell cultivation method based on the hematopoietic stem cell culture medium. The hematopoietic stem cell culture medium is made from amino acids, vitamins, salts, fats, phaseolin polypeptide and cytokines, with one or any of trace elements, organic small molecules, hematopoietic stem cell proliferator, plant extract and animal extract; the hematopoietic stem cell culture medium can quickly proliferate CD34+ cells different in species, and proliferating speed and telomere length of the CD34+ cells is evidently increased; hematopoietic stem cells cultivated via the hematopoietic stem cell culture medium have constant good cell functions, the hematopoietic stem cell culture medium has the functions, including, but not limited to, anticancer action, anti-ageing action and life prolonging, and has promising application prospect.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a hematopoietic stem cell culture medium, its application and a stem cell culture method. Background technique [0002] Many studies have shown that hematopoietic stem cells play a key role in body fluid circulation: on the one hand, red blood cells and lymphocytes in body fluids are differentiated from hematopoietic stem cells. However, under normal physiological conditions, hematopoietic stem cells mainly exist in the bone marrow. Released into peripheral blood after treatment; on the other hand, cell renewal in body fluids also depends on the number and differentiation potential of hematopoietic stem cells. With the rise of personalized regenerative medicine technology, hematopoietic stem cells play an increasingly important role in anti-aging and disease treatment (such as diabetes, tumors, immune system diseases and ischemic tissue necrosis, etc.). [0003] With the rise of pers...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789
CPCA61K35/28C12N5/0647C12N2500/76C12N2501/105C12N2501/11C12N2501/113C12N2501/115C12N2501/125C12N2501/135C12N2501/14C12N2501/145C12N2501/165C12N2501/19C12N2501/22C12N2501/2302C12N2501/235C12N2501/24C12N2501/25C12N5/0018C12N2500/33C12N2500/36C12N2500/38
Inventor 刘小青
Owner SHENZHEN FULIXIN HEALTH IND DEV LTD
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