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Process and model for separating senescence cells

A technology of cells and cell groups, applied in the fields of biotechnology and cell biology, can solve the problems of small proportion and difficulties in the research of aging cells

Inactive Publication Date: 2013-02-06
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, due to the morphological similarity between mother cells and daughter cells and the small proportion of senescent cells in the entire cell population, the isolation of senescent yeast cells requires complex procedures and special devices (Park, P.U. et al. (2002), Separation of mother and daughter cells.Methods Enzymol351, 468-477), it is very difficult to obtain a large number of senescent cells for research on a high-throughput level, and the isolation of senescent cells has thus become a bottleneck and restrictive factor in research in this field

Method used

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  • Process and model for separating senescence cells
  • Process and model for separating senescence cells
  • Process and model for separating senescence cells

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0115] Embodiment 1 is in the separation method of the cell of same age stage

[0116] 1. Strains and culture conditions

[0117] The strain used in the experiment was Candida albicans (Candida albicans) wild-type strain SC5314 (purchased from ATCC).

[0118] Yeast-like cells were cultured at 30°C in YPD medium. Under this culture condition, the growth state of Candida albicans is as follows: figure 1 A is shown on the left.

[0119] The culture condition of hyphal cells is 37°C, YPD+15% calf serum. Under this culture condition, the growth state of Candida albicans is as follows: figure 1 As shown in the figure in A. When the mycelial cells of Candida albicans are returned from this culture condition to the culture condition of 30°C and the YPD medium is continued to culture, it can produce yeast-like daughter cells such as figure 1 A is shown on the right.

[0120] The formula of YPD medium is as follows:

[0121] Peptone 2%,

[0122] Yeast Extract 1%,

[0123] Gluc...

Embodiment 2

[0137] Example 2. Oxidative damage proteins are continuously accumulated during the aging process of Candida albicans

[0138] The resistance of aging cells to certain stresses such as oxidative stress will be greatly reduced, so the accumulation of oxidatively damaged proteins in aging cells should also be more than that in young cells. A carbonyl group will be irreversibly introduced into the protein molecule oxidized and modified. The inventors used the chemical characteristics of the carbonyl group and combined with the method of immunoassay to analyze the levels of oxidatively damaged proteins in the isolated Candida albicans cells of different ages. comparative analysis.

[0139] The present inventors respectively took Candida albicans cell populations with an average lifespan of 22.9, 13.1, 8.4, and 5.4 generations, and carried out oxidative damage protein detection experiments.

[0140] Oxidative damage protein detection experiments are as follows:

[0141] The cells...

Embodiment 3

[0143] Example 3. Glycogen levels increase continuously along with the senescence process of Candida albicans cells

[0144] Senescent cells are cells that are not metabolically active, so certain metabolites accumulate in senescent cells. The process of cell aging in yeast is accompanied by gluconeogenesis and energy storage, so the level of glycogen in aging cells is higher (Lin, S.S. et al. (2001), Enhanced gluconeogenesis and increased energy storage as hallmarks of aging in Saccharomyces cerevisiae. J BiolChem276, 36000-36007.). Genomic studies have shown that glycogen synthesis genes are upregulated in aging cells (Lesur, I. et al. (2004), The transcriptome of prematurely aging yeast cells is similar to that of telomerase-deficient cells. Mol BiolCell 15, 1297-1312. ). In higher organisms, glycogen accumulation occurs not only during physiological aging, but also under some pathological conditions.

[0145] The present inventors took Candida albicans cell populations ...

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Abstract

An isolated cell population of Candida is provided, as well as the method for preparing the cell population and its uses, in which most cells of the cell population are at the same age stage. Based on the feature that candida cells have two distinct growth forms during different growth conditions, methods for preparing cell population at the same age stage on a large scale have been developed. The cell population at the same age stage may be used to study cells growth, development and the characterizations of cells at various age stages, as well as cellular aging.

Description

technical field [0001] The present invention belongs to the fields of biotechnology and cell biology; more specifically, the present invention relates to a method for producing cell groups at the same age and cell groups obtained by the method. Background technique [0002] Aging is one of the hot issues that have been widely concerned and researched for a long time. It usually refers to the gradual, genetic factors, complex morphological structure and irreversible degenerative changes in physiological functions that occur with age. The main manifestations are Declining fertility and rising mortality. Aging is relatively conservative in evolution, and there is a physiological process of aging from the highest organism—human beings to very low prokaryotes—bacteria. [0003] Cellular senescence is the basic unit of biological aging, and it is a process of complex physiological and biochemical changes in cells, which may eventually be reflected in the morphology, structure and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20
CPCC12N1/16
Inventor 周金秋付晓虹孟飞龙
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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