Characteristics of morphology,stem cell related markers and metastasis ability of colorectal cancer cells in vitro
Author(s): JIA Jian-feng, HE Chang-lin
Pages: 290-293
Year: 2014 Issue: z1
Journal: Journal of Laboratory Medicine and Clinical Sciences
Keyword: colorectal carcinoma cell lines; cancer stem/tumor-initiating cells; stem cell related marker; serum-free culture; CD133;
Abstract: Objective To explore the changes of cell morphology,expression of stem cell related markers and metastasis ability of colorectal cancer stem cells after differentiation in vitro.Methods Tumor stem cells of the line CD133+ were obtained from colorectal cancer cell line HCT116 cells ,serum-free culture isolate CD133 + cells,and serum levels of differentiation,under phase contrast microscope to observe the morphological changesCD133+ cell were isolated by serum-free culture and then cultured to differentiate in medium containing 10% fetal bovine serum. The morphology of the cells was observed under phase contrast microscope.Cells were collected respectively before differentiation,after differentiate 7,14 days .The cell surface markers such as CD133,N-cadherin and Vimentin were investigated by flow cytometry,RT-PCR and immunofluorescence .Cell metastasis assay was applied to examine cell metastatic ability in vitro.Results (1)HCT116 cells were formed cancer cell spheres in SFM,Sphere cells were able to produce a heterogeneous cell population that could produce spheres.Cancer cell spheres were induced to differenti-ate by serum supplemented medium,these differentiated cell were adherence growth and kept the same morphology with the parental cells.(2)These cancer cell spheres were founded expressing markers such as CD133+,N-cadherin and Vimentin was higher than differentiated cells.The expression rates of CD133 was 20% after seven days ,90%after forteen days,and 0.5% of the differentiated cells.(3)The migrated cells number of CD133+ cells were 104.67 ±14.64,which were significant higher than differentiated cells 28.67±6.5(P <0.05).Conclusion The changes of morphology,cell marker expression and metastasis ability in the high-expression CD133+ CRC cells are oriented dif-ferentiated into the same origin CRC cells,and the metastasis ability of differentiated cells is higher than the cell sphere.
Pages: 290-293
Year: 2014 Issue: z1
Journal: Journal of Laboratory Medicine and Clinical Sciences
Keyword: colorectal carcinoma cell lines; cancer stem/tumor-initiating cells; stem cell related marker; serum-free culture; CD133;
Abstract: Objective To explore the changes of cell morphology,expression of stem cell related markers and metastasis ability of colorectal cancer stem cells after differentiation in vitro.Methods Tumor stem cells of the line CD133+ were obtained from colorectal cancer cell line HCT116 cells ,serum-free culture isolate CD133 + cells,and serum levels of differentiation,under phase contrast microscope to observe the morphological changesCD133+ cell were isolated by serum-free culture and then cultured to differentiate in medium containing 10% fetal bovine serum. The morphology of the cells was observed under phase contrast microscope.Cells were collected respectively before differentiation,after differentiate 7,14 days .The cell surface markers such as CD133,N-cadherin and Vimentin were investigated by flow cytometry,RT-PCR and immunofluorescence .Cell metastasis assay was applied to examine cell metastatic ability in vitro.Results (1)HCT116 cells were formed cancer cell spheres in SFM,Sphere cells were able to produce a heterogeneous cell population that could produce spheres.Cancer cell spheres were induced to differenti-ate by serum supplemented medium,these differentiated cell were adherence growth and kept the same morphology with the parental cells.(2)These cancer cell spheres were founded expressing markers such as CD133+,N-cadherin and Vimentin was higher than differentiated cells.The expression rates of CD133 was 20% after seven days ,90%after forteen days,and 0.5% of the differentiated cells.(3)The migrated cells number of CD133+ cells were 104.67 ±14.64,which were significant higher than differentiated cells 28.67±6.5(P <0.05).Conclusion The changes of morphology,cell marker expression and metastasis ability in the high-expression CD133+ CRC cells are oriented dif-ferentiated into the same origin CRC cells,and the metastasis ability of differentiated cells is higher than the cell sphere.
Related Articles