Method for enriching tumor stem cells through continuous low dose radiation
A tumor stem cell, low-dose technology, applied in the biological field, can solve the problems of difficult sample acquisition and small proportion of tumor stem cells.
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Embodiment 1
[0028] Example 1 Enrichment of MDA-MB-231 tumor stem cells
[0029] The breast cancer cell line MDA-MB-231 was cultured under routine laboratory conditions, after isotope 60 Co radiation source, 2Gy irradiation treatment (1:17min at a dose of 139.09cGy / min), and routine culture for 1-2 weeks after treatment, when the cell growth condition is stable, accept again 60 Co and 2Gy irradiation treatments were repeated, and cells (n*2Gy) irradiated with 1*2Gy, 2*2Gy and 3*2Gy were collected respectively.
Embodiment 2
[0030] Example 2 Enrichment of MCF7 tumor stem cells
[0031] The breast cancer cell line MCF7 was cultured under routine laboratory conditions, after isotope 60 Co radiation source, 2Gy irradiation treatment (1:17min at a dose of 139.09cGy / min), and routine culture for 1-2 weeks after treatment, when the cell growth condition is stable, accept again 60 Co and 2Gy irradiation treatments were repeated, and cells (n*2Gy) irradiated with 1*2Gy, 2*2Gy and 3*2Gy were collected respectively.
Embodiment 3
[0032] Example 3 Flow cytometry analysis of enrichment of tumor stem cells
[0033] The cells obtained in Examples 1 and 2 were incubated with two antibodies, CD24-APC and CD44-FITC, respectively, and flow cytometry analysis found that continuous low-dose radiation in the two cell lines could transform CD44 + CD24 - / low Especially in the MCF7 cell line, the proportion of tumor stem cells increased by nearly 10 times, thus indicating that the use of continuous low-dose radiation breast cancer cell lines can increase the proportion of tumor stem cells (see figure 1 ).
[0034] The above-mentioned flow cytometry experiment process is briefly described as follows: using the method of flow cytometry sorting, firstly collect different batches of processed cells, wash them with phosphate buffered solution PBS, and then use two kinds of CD24-APC and CD44-FITC The cells were incubated with the antibody, and 635nm and 488nm excitation light were selected respectively, and the quadrant...
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