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Application of baicalein in promoting the expansion of breast stem cells in vitro and maintaining their stem cell characteristics

A mammary gland stem cell and in vitro expansion technology is applied in the field of traditional Chinese medicine extracts that promote in vitro culture and expansion of mammary gland stem cells and maintain their stem cell characteristics, and can solve the problems of baicalin mammary stem cells that have not yet been seen.

Active Publication Date: 2021-02-19
SHANGHAI HENGSAI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] However, there is no report on the use of baicalin in expanding breast stem cells in vitro

Method used

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  • Application of baicalein in promoting the expansion of breast stem cells in vitro and maintaining their stem cell characteristics
  • Application of baicalein in promoting the expansion of breast stem cells in vitro and maintaining their stem cell characteristics
  • Application of baicalein in promoting the expansion of breast stem cells in vitro and maintaining their stem cell characteristics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] [Example 1] Baicalin can promote the in vitro expansion of breast stem cells

[0038] 1. Single-cell preparation: After the adult mice were sacrificed, the fourth pair of mammary gland tissues were taken out, and the shredded mammary glands were digested by collagenase. The digestion time was about 2 hours. The digestive juice was RPMI-1640, 0.01M Hepes, 5% FBS, collagenase 3 was added to the digestion solution, and the addition amount was 1.5mg / mL; red blood cell lysate was added to the digested mammary gland to remove red blood cells, and the red blood cell lysis time was about 5 minutes; Cells, trypsinization time is about 10 minutes, add deoxyribonuclease 1 to trypsin to remove DNA;

[0039] 2. Antibody staining: Add the following antibodies to the prepared single cells: anti-Ter119-FITC, anti-CD31-FITC, anti-CD45-FITC, anti-CD29-APC and anti-CD24-PE-Cy7 for staining, The antibody concentration is 1:200, and the staining time is 20 minutes;

[0040] 3. Flow cytome...

Embodiment 2

[0043] [Example 2] Baicalin promotes the in vivo reconstruction ability of breast basal stem cells

[0044] Transplant mammary stem cells of the 5th passage into the mammary fat pad of 3-week-old nude mice, and check the growth of mammary glands after nude mice were cultured for 4 to 8 weeks to determine the regenerative ability of mammary stem cells. Transplantation experiments were carried out on the cell clones after serial passage to investigate the probability of mammary gland tissue regeneration.

[0045] Mammary gland basal stem cells are cultured in 3D in vitro in Matri-Gel and continuously passaged. After continuous passage, basal stem cells will gradually differentiate and lose their stem cell characteristics, and lose their ability to regenerate mammary glands. The present invention finds that adding Baicalin treatment during the continuous passage can make the mammary gland basal stem cells continue to maintain the characteristics of stem cells after the continuous...

Embodiment 3

[0046] [Example 3] Baicalin promotes the development of mammary glands:

[0047] Get the mice of 3 weeks, take the mode of gavage to carry out Baicalin processing, the dose of Baicalin processing is 200mg / Kg, get the mice after Baicalin processing two weeks and (5 weeks mice) three weeks (6 weeks mice) respectively , the fourth pair of mammary glands was stained with Carmine, compared with the control group to investigate the extension and side branching of mammary ducts after Baicalin treatment. Compared with the control group, it was found that the development of mammary glands in mice was significantly accelerated after gavage with Baicalin, and mammary ducts extended faster and had more side branches (such as Figure 4 shown).

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Abstract

The invention discloses application of baicalin to the promotion of in vitro expansion of mammary stem cells and the maintenance of stem cell characteristics. As discovered by the invention, the baicalin can promote clone formation and self-renewal of the mammary stem cells during in vitro culture; the baicalin can improve the reconstruction efficiency of the mammary stem cells during in vivo mammary gland reconstruction, and can promote the development of the mammary gland when mice eat the baicalin at puberty; the baicalin can directly up-regulate the expression of a molecular marker proteinC receptor on the surface of the mammary stem cells. Thus, the baicalin can realize in vitro culture and expansion of the mammary stem cells instead of Wnt3A proteins or hormone. In vivo transplantation experiments also show that these mammary stem cells can still keep developing potential even after in vitro long-term continuous cell culture, and can develop into breast organs with complete functions and normal forms.

Description

technical field [0001] The invention belongs to the technical field of in vitro expansion of stem cells, and more specifically relates to a traditional Chinese medicine extract and a method for promoting the in vitro culture and expansion of breast stem cells and maintaining the characteristics of stem cells. Background technique [0002] Stem cells are a class of primitive cells capable of self-renewal and multipotent differentiation potential. Stem cells are currently divided into three major categories: embryonic stem cells with totipotency, adult stem cells with pluripotency, and induced pluripotent stem cells (iPSC) produced by induction. kind. With the continuous progress in the field of stem cell research, more and more different types of stem cells have shown the potential for clinical application, such as mesenchymal stem cells for the treatment of osteoarthritis, iPSC can be directed to differentiate into hematopoietic stem cells for clinical treatment of blood dis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0631C12N2501/11C12N2501/999C12N2513/00C12N2533/90
Inventor 丁衡
Owner SHANGHAI HENGSAI BIOLOGICAL TECH CO LTD
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