Selective massive purification amplification method of stem cells

A stem cell and selective technology, applied in the field of biomedicine, can solve the problems of stem cell stemness loss, abnormal cell proliferation signal, difficulty in simulating the stem cell microenvironment, etc., and achieve the effect of large-scale expansion.

Active Publication Date: 2015-11-11
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is difficult to simulate the microenvironment of stem cells in vivo under general cell culture conditions in vitro, which often leads to abnormal cell proliferation signals, resulting in the loss of stem cell stemness.

Method used

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  • Selective massive purification amplification method of stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1. Extract single-strand fibers from the medical degreasing surface, and the fiber diameter is 5-10 μm.

[0022] 2. Plasma surface treatment of absorbent cotton fibers (PDC-M plasma cleaning machine). The conditions of ammonia plasma treatment were: input power 20W; treatment time 60s; activation frequency 13.65MHz; pressure 15pa; ammonia gas flow rate 150mL / min.

[0023] 3. After the surface treatment of the defatted fiber is completed, use ultraviolet light to irradiate for 10 hours to sterilize for later use.

[0024] 4. Use 1% agarose (Ultrapureagarose, Invitrogen, Carlsbad, CA) to coat the bottom of the culture dish, and then inoculate the PC12 cell suspension, for example, inoculate 2×10 cells in each well of a six-well plate. 4 cells / mL.

[0025] 5. Put the sterilized degreased fibers in a petri dish and culture them for 5 days. During the culture period, replace the culture medium every 2-3 days. Finally, collect the cell balls on the fibers. The diameter of t...

Embodiment 2

[0028] Basically the same as Example 1, the difference is that the anti-adsorption cell material used is gelatin, and the treatment conditions of ammonia plasma are: treatment time is 10s; frequency is 10kHz; pressure is 10pa; ammonia gas flow rate is 50mL / min, After the surface treatment of the defatted fiber is completed, it is sterilized by high temperature and high pressure; the present invention cultures adipose-derived mesenchymal stem cells for 9 days, and the number ranges from 2×10 4 Cells / mL expanded to 7×10 5 cells / mL, the expressions of Oct4, Sox2 and Nanog, which represent the characteristics of stem cells, also increased significantly.

Embodiment 3

[0030] Basically the same as Example 1, the difference is that the treatment conditions of ammonia plasma are: treatment time is 35s; frequency is 40kHz; Bacteria; After the surface treatment of defatted fiber is completed, it is soaked and sterilized by iodine solution, and the present invention cultivates induced pluripotent stem cells for 15 days, and the number is from 2×10 4 Cells / mL expanded to 3×10 6 cells / mL, the expressions of Oct4, Sox2 and Nanog, which represent the characteristics of stem cells, also increased significantly.

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Abstract

The invention discloses a selective massive purification amplification method of stem cells. The method comprises the following step: carrying out selective amplification on stem cells by using absorbent cotton fibers or modified absorbent cotton fibers. The absorbent-cotton-fiber-based high-polymer material is utilized to effectively enhance and purify the stem cells and keep the stem cell characteristics unchanged, thereby being beneficial to implementing abundant amplification of the stem cells.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for selectively purifying and expanding stem cells in large quantities based on absorbent cotton fibers. The use of absorbent cotton fibers can effectively increase the value of stem cells and maintain the characteristics of stem cells, which is conducive to the realization of a large number of stem cells. Background technique [0002] Stem cells play an important role in life science research, medical treatment and drug screening. However, the growth conditions of stem cells are harsh, the number in the organism is rare, and they are prone to aging and differentiation under general culture conditions. Therefore, how to obtain a large number of stem cells is one of the difficult problems in the field of life sciences. [0003] In addition to hematopoietic stem cells, other types of stem cells generally require adherent growth or aggregate growth (neural stem cells) to achiev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/095C12N5/071
Inventor 肖忠党孙博王秀王俊华胡飞虎
Owner SOUTHEAST UNIV
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