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Culture system and method for amplifying hematopoietic stem cells and/or hematopoietic progenitor cells, hematopoietic stem cells, and hematopoietic progenitor cells

A technology of hematopoietic stem cells and hematopoietic progenitor cells, which is applied in the fields of biotechnology and medicine, can solve the problems of insufficient single signaling pathway and limit the scale of clinical application, and achieve the effects of safe clinical use, improved in vivo reconstruction effect, and obvious amplification effect

Active Publication Date: 2018-06-29
NEWISH TECH (BEIJING) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, small molecular compounds have a wide range of targets, and their signaling pathways are not single enough, which may limit the scale of their clinical application

Method used

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  • Culture system and method for amplifying hematopoietic stem cells and/or hematopoietic progenitor cells, hematopoietic stem cells, and hematopoietic progenitor cells
  • Culture system and method for amplifying hematopoietic stem cells and/or hematopoietic progenitor cells, hematopoietic stem cells, and hematopoietic progenitor cells
  • Culture system and method for amplifying hematopoietic stem cells and/or hematopoietic progenitor cells, hematopoietic stem cells, and hematopoietic progenitor cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] In this example, the culture system provided by the present invention is used to culture CD34+ cells.

[0068] Cultivate CD34+ cells with reference to the culture conditions in Table 1, divide them into experimental group (chemical small molecule is SP600125) and control group (use DMSO instead of chemical small molecule, DMSO:medium volume ratio is 1:10000), place at 37°C, 5 In the cell culture incubator with % carbon dioxide concentration, change the medium in half every 2 days.

[0069] After culturing until the 7th day, the ratio of CD34+CD45RA- cells was detected and counted to calculate the amplification factor. The result is as figure 1 As shown, the amplification factor of CD34+CD45RA- in the experimental group was about 15 times, and the amplification factor of CD34+CD45RA- in the control group was about 5 times.

[0070] On the 7th day of culture, in vivo transplantation experiments were carried out at the same time. The expanded CD34+ cells of the experimen...

Embodiment 2

[0073] In this example, the culture system provided by the present invention is used to culture CD34+ cells.

[0074] Cultivate CD34+ cells with reference to the culture conditions in Table 1, divide them into experimental group (chemical small molecule is SP600125) and control group (use DMSO instead of chemical small molecule, DMSO:medium volume ratio is 1:10000), place at 37°C, 5 In the cell culture incubator with % carbon dioxide concentration, change the medium in half every 2 days.

[0075] After culturing until the 7th day, the ratio of CD34+CD45RA- cells was detected and counted to calculate the amplification factor. The result is as Figure 4 As shown, the expansion of CD34+CD45RA- cells in the experimental group was 2.5 times that of the CD34+CD45RA- cells in the control group.

[0076] On the 7th day of culture, the in vivo transplantation experiment was carried out, and the expanded CD34+ cells of the experimental group and the control group were inoculated into ...

Embodiment 3

[0079] In this example, the culture system provided by the present invention is used to culture CD34+ cells.

[0080] Cultivate CD34+ cells with reference to the culture conditions in Table 1, divide them into experimental group (chemical small molecule is SP600125) and control group (use DMSO instead of chemical small molecule, DMSO:medium volume ratio is 1:10000), place at 37°C, 5 In the cell culture incubator with % carbon dioxide concentration, change the medium in half every 2 days.

[0081] After culturing until the 7th day, the ratio of CD34+CD45RA- cells was detected and counted to calculate the amplification factor. see results Figure 7 , wherein the expansion of CD34+CD45RA- cells in the experimental group is twice the number of CD34+CD45RA- cells in the control group.

[0082] On the 7th day of culture, the in vivo transplantation experiment was carried out, and the expanded CD34+ cells of the experimental group and the control group were inoculated into immunode...

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Abstract

The present invention relates to a culture system and method for amplifying hematopoietic stem cells and / or hematopoietic progenitor cells, hematopoietic stem cells, and hematopoietic progenitor cells. The culture system comprises: a basal medium suitable for stem cell expansion; and a JNK signaling pathway inhibitor. The culture system provided by the invention can expand hematopoietic stem cellsin vitro, and the amplification effect thereof is obvious, and the CD34+CD45RA-labeled hematopoietic stem cells can be amplified nearly 100 times.

Description

technical field [0001] The invention mainly relates to the fields of biotechnology and medicine, and in particular to a culture system and method for expanding hematopoietic stem cells and / or hematopoietic progenitor cells, hematopoietic stem cells and hematopoietic progenitor cells. Background technique [0002] Hematopoietic stem cells are a type of blood stem cells that have self-renewal ability and multi-lineage differentiation potential and can complete long-term (at least 4 months) hematopoietic reconstruction in vivo. Compared with hematopoietic stem cells, hematopoietic progenitor cells have weaker reconstruction ability and cannot maintain the level of reconstruction in vivo for a long time. Generally, within 2 months, the reconstruction cells will gradually lose. In 1961, scientists first demonstrated the existence of hematopoietic stem cells using the method of spleen nodules in mice. Later, Weissman and other laboratories in the United States expanded people's u...

Claims

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Application Information

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IPC IPC(8): C12N5/0789
CPCC12N5/0647C12N2501/999
Inventor 罗小霞
Owner NEWISH TECH (BEIJING) CO LTD
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