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Optimization of culture medium for the isolation and propagation of human breast cancer cells from primary tumour biopsies

http://repository.vnu.edu.vn/handle/VNU_123/32547

Breast cancer cells from patients hold an important role in antigen production for immunotherapy, drug testing, and cancer stem cell studies.

To date, although many studies have been conducted to develop protocols for the isolation and culture of breast cancer cells from tumour biopsies, the efficiencies of these protocols remain low.

This study aimed to identify a suitable medium for the isolation and propagation of primary breast cancer cells from breast tumour biopsies.

Breast tumour biopsies were obtained from hospitals after all patients had given their written informed consent and were cultured according to the expanding tumour method in 3 different media: DMEM/F12 (Dulbecco’s Modified Eagle Medium: Nutrient Mixture F-12) supplemented with 10% FBS (Fetal bovine serum) and 1% antibiotic-antimycotic (Medium D); Medium 171 supplemented with 1X MEGS (Mammary Epithelial Growth Supplement) and 1% antibiotic-antimycotic (Medium M); or a 1:1 mixture of Medium D and Medium M (Medium DB).

The cell culture efficiency was evaluated by several criteria, including the time of cell appearance, cell morphology, capability of proliferation, cell surface marker expression, ALDH (Aldehyde dehydrogenases) activity, karyotype, and tumour formation capacity in immune-deficient mice.

Notably, primary cancer cells cultured in Medium DB showed a high expression of breast cancer stem cell surface markers (including CD44(+)CD24(-) and CD49f(+)), low expression of stromal cell surface markers (CD90), high ALDH activity, an abnormal karyotype, and high tumour formation capacity in immune-deficient mice.

These findings suggested that Medium DB was suitable to support the survival and proliferation of primary breast cancer cells as well as to enrich breast cancer stem cells.

 

Title: Optimization of culture medium for the isolation and propagation of human breast cancer cells from primary tumour biopsies
Authors: Vu, Thanh Binh
Le, Thi Hanh
Phan, Chinh Nhan Lu
Keywords: Breast cancer cell
breast cancer stem cell
culture medium
primary cancer cell
Issue Date: 2015
Publisher: BIOMEDPRESS, BIOMEDPRESS, HO CHI MINH, 00000, VIETNAM
Citation: ISIKNOWLEDGE
Abstract: Breast cancer cells from patients hold an important role in antigen production for immunotherapy, drug testing, and cancer stem cell studies. To date, although many studies have been conducted to develop protocols for the isolation and culture of breast cancer cells from tumour biopsies, the efficiencies of these protocols remain low. This study aimed to identify a suitable medium for the isolation and propagation of primary breast cancer cells from breast tumour biopsies. Breast tumour biopsies were obtained from hospitals after all patients had given their written informed consent and were cultured according to the expanding tumour method in 3 different media: DMEM/F12 (Dulbecco’s Modified Eagle Medium: Nutrient Mixture F-12) supplemented with 10% FBS (Fetal bovine serum) and 1% antibiotic-antimycotic (Medium D); Medium 171 supplemented with 1X MEGS (Mammary Epithelial Growth Supplement) and 1% antibiotic-antimycotic (Medium M); or a 1:1 mixture of Medium D and Medium M (Medium DB). The cell culture efficiency was evaluated by several criteria, including the time of cell appearance, cell morphology, capability of proliferation, cell surface marker expression, ALDH (Aldehyde dehydrogenases) activity, karyotype, and tumour formation capacity in immune-deficient mice. Notably, primary cancer cells cultured in Medium DB showed a high expression of breast cancer stem cell surface markers (including CD44(+)CD24(-) and CD49f(+)), low expression of stromal cell surface markers (CD90), high ALDH activity, an abnormal karyotype, and high tumour formation capacity in immune-deficient mice. These findings suggested that Medium DB was suitable to support the survival and proliferation of primary breast cancer cells as well as to enrich breast cancer stem cells.
Description: BIOMEDICAL RESEARCH AND THERAPY Volume: 2 Issue: 2 Pages: 207-219 Published: JAN 2015 ; TNS05790
URI: http://repository.vnu.edu.vn/handle/VNU_123/32547
ISSN: 2198-4093
Appears in Collections: Bài báo của ĐHQGHN trong Web of Science

 

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