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A cell culture medium and its application in culturing megakaryotic progenitor cells

A cell culture and stem cell technology, applied in the field of cell culture, can solve the problems of slow proliferation rate and small number of megakaryotic progenitor cells, and achieve the effect of promoting growth and proliferation and reducing risks.

Active Publication Date: 2019-02-22
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the current method for culturing megakaryotic progenitor cells in vitro proliferates slowly, and the number of megakaryotic progenitor cells contained in the obtained cells is not large. Therefore, it is necessary to research and develop methods for high-yield and high-purity megakaryotic progenitor cells.

Method used

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  • A cell culture medium and its application in culturing megakaryotic progenitor cells
  • A cell culture medium and its application in culturing megakaryotic progenitor cells
  • A cell culture medium and its application in culturing megakaryotic progenitor cells

Examples

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Embodiment 1～3

[0051] The composition of the medium is shown in Table 1:

[0052] Table 1 Examples 1 to 3 of medium components

[0053]

[0054] 1. Take 20ml of cord blood, add an equal volume of normal saline to dilute, add the diluent to one-third of the volume of Ficoll separation solution, centrifuge at 700g for 20-30min, the speed of rise and fall will drop to zero. Absorb the middle albuginea layer;

[0055] 2. Wash the cells in the albuginea layer twice with physiological saline, add the medium of Examples 1 to 3 according to the counting results, and adjust the inoculation density to 1×10 respectively. 5 cell / ml, put 5% CO 2 , Cultivate in 37℃ incubator.

[0056] 3. Aspirate the suspended cells after 2 hours and place them in culture flasks containing fresh culture medium (the culture medium of Examples 1 to 3);

[0057] 4. After that, change the medium in half every 3 days and keep the cell density at 1×10 5 cell / ml;

[0058] 5. Count the cells cultured for 14 days and 20 days.

[0059] Each t...

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Abstract

The invention relates to the field of cell culture, in particular to a cell culture solution and application thereof to culture of megakaryocyte progenitor cells. According to the invention, WNT3a protein, insulin, Flt-3L, albumin, non-essential amino acid, thrombopoietin, stem cell growth factors, interleukin 3 and interleukin 6 are compounded, and a basal culture medium is added, so that richer nutrition can be provided for cells. Experiments show that the culture solution provided by the invention is used for culture of megakaryocyte progenitor cells and can selectively promote growth and proliferation of the megakaryocyte progenitor cells. After the culture solution provided by the invention cultures the megakaryocyte progenitor cells for 20 days, the number of cells is increased by 400-450 times, and the CD34+ and CD41+ double-positive expression rate in the cells is not lower than 35%. The effect is remarkably superior to that of a culture medium in the prior art (p is less than 0.01).

Description

Technical field [0001] The invention relates to the field of cell culture, in particular to a cell culture fluid and its application in culturing megakaryotic progenitor cells. Background technique [0002] Megakaryocyte progenitor cell (megakaryocyte progenitor cell) is a cell that enables people to produce megakaryocytes. It is derived from bone marrow progenitor (CFU-GEMM), referred to as megakaryocyte progenitor cell. Megakaryotic progenitor cells exist in the IL-6R-cell population, and CD34+ cells can be divided into two cell subgroups, expressing IL-6R and not expressing IL-6R. IL-6R+ cells can be stimulated to form granulo-macrophages, Lymphocytes and granulocytes, while IL-6R-cells can form erythroid and megakaryocytes when given IL-6 / sIL-6R. Hematopoietic stem cells first differentiate into megakaryocyte progenitor cells, also called colony forming unit-megakaryocyte (CFU-Meg). After megakaryocyte progenitor cells differentiate into mature megakaryocytes, their edges r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/078
Inventor 陈海佳王一飞葛啸虎万桦
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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