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Method for monitoring number of cell division, and application thereof

A technology of cell division and number of divisions, applied in biochemical equipment and methods, determination/inspection of microorganisms, etc., can solve the problems of large animals and time, inaccuracy, etc., to achieve the effect of simple operation, accurate and reliable results

Active Publication Date: 2011-10-19
BEIJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Not only does it require a lot of animals and time, it's not precise enough

Method used

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  • Method for monitoring number of cell division, and application thereof
  • Method for monitoring number of cell division, and application thereof
  • Method for monitoring number of cell division, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1, the method for monitoring the number of cell divisions in vitro

[0063] 1. Materials:

[0064] 1. BrdU (Sigma-Aldrich, St. Louis, MO)

[0065] 2. Mouse IgG against BrdU (Becton Dickinson, San Jose, CA)

[0066] 3. FITC-coupled donkey anti-mouse IgG (Jackson ImmunoResearch, West Grove, PA)

[0067] 4. DAPI (Sigma-Aldrich, St. Louis, MO)

[0068] 5. PI (Sigma-Aldrich, St. Louis, MO)

[0069] 6. o-Phenylenediamine (Sigma-Aldrich, St.Louis, MO)

[0070] 7. Dulbecco's modified Eagle's medium (DMEM, Gibco, Long Island, NY)

[0071] 8. Fetal Bovine Serum (Gibco, New Zealand)

[0072] 9. The CHO cell line was purchased from ATCC, the product catalog number is CCL-61.

[0073] 10. Paraformaldehyde (Sigma-Aldrich, St. Louis, MO)

[0074] 2. The theoretical number of divisions of CHO cells at each monitoring time point

[0075] The CHO cell cycle is about 12 hours; the total chromosome number of CHO cells is 19 (N=19).

[0076] The cells to be detected wer...

Embodiment 2

[0177] Embodiment 2, the method for monitoring the number of cell divisions in vivo

[0178] Material:

[0179] 1. BrdU (Sigma-Aldrich, St. Louis, MO)

[0180] 2. Mouse IgG against BrdU (Becton Dickinson, San Jose, CA)

[0181] 3. FITC-coupled donkey anti-mouse IgG (Jackson ImmunoResearch, West Grove, PA)

[0182] 4. DAPI (Sigma-Aldrich, St. Louis, MO)

[0183] 5. n-propyl gallate (Sigma-Aldrich, St.Louis, MO)

[0184] 6. SD strain rats (Vital River Laboratories, Beijing)

[0185] 7. Paraformaldehyde (Sigma-Aldrich, St. Louis, MO)

[0186] 8. Polylysine (Sigma-Aldrich, St. Louis, MO)

[0187] 9. Cryostat (Leica 3050S)

[0188]Liver regeneration background: It is known that almost all liver cells are in the non-proliferative quiescent state (G0 phase) of the cell cycle under normal conditions. Compensatory proliferation will start after liver resection. Liver regeneration in rats lasts for 7 days, and then the liver regenerates Returning to their pre-surgery weight, the...

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Abstract

The invention discloses a method for monitoring number of cell division, and an application thereof. The method for assistant monitoring the number of the cell division comprises the following steps: (1) adopting base analogs for marking a cell to be monitored, wherein a single-stranded DNA of all double-stranded DNA of the marked cell to be monitored is marked with the base analog after marking,the marked cell to be monitored is recorded as a undivided initial cell, a base analog marked chromosome is recorded as a marked chromosome; (2) determining division number of the initial cell according to the following method 1) or the method 2): 1) observing the marked chromosome in the daughter cell generated from the division of the initial cell, then observing and counting the amount of the marked chromosome in single daughter cell if each cell of the daughter cell contains the marked chromosome, and obtaining the division number of the initial cell according to the amount of the marked chromosome in the single daughter cell. The method provides correct and reliable result, and does not require extra operations during tracking except routine cell culture, routine animal feeding and final cell dyeing operation. Therefore, the method provided by the present invention has advantages of simple operation, and time saving and labour saving.

Description

technical field [0001] The invention relates to a method for monitoring the number of cell divisions and its application. Background technique [0002] It has wide application and significance for effective and precise detection of cell proliferation speed. For example, in the screening of anticancer drugs, when tumor cells cultured in vitro are used, techniques such as MTT cell counting and living cell microscopy counting are usually used to count the number of cells at different time points, and then make a change curve to judge the proliferation The speed changes quickly and slowly. When performing in vivo tumorigenesis assays, it is not possible to perform multiple cell counts on the same tumor nodule. Existing methods mainly measure the diameter and number of tumor nodules, and estimate the growth volume change of the tumor through the statistics of a large number of parallel samples, so as to reflect the growth speed change of tumor cells. Not only does it require a...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06C12Q1/18
Inventor 何大澄吴艺舟
Owner BEIJING NORMAL UNIVERSITY
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