I am working on colon cancer stem cells to see the impact of combinational therapy.
Dear Aditi!
Please You look at this article:
Article Colorectal Cancer Migration and Invasion Initiated by microRNA-106a
Cell Lines
The immortalized human colon epithelial cell line,NCM640, was a gift from J.Q. Zhang and cultured in DMEM supplemented with 10% FBS HT29, SW480, LOVO, SW1116, and SW480 cell lines were purchased from the ATCC (Manassas, VA, USA) and cultured under conditions in accordance with the manufacturer’s instructions.
In vitro Migration and Invasion Assays
The migration and invasion potential of CRC cells was evaluated, as described previously with slight modifications [34].
CRC cells (2.5×105) were plated on uncoated upper chambers (24-well inserts; pore size, 8 µm; BD Bioscience, Bedford, MA, USA) for transwell migration assays. CRC cells (2.5×105) were placed on Matrigel-coated upper chambers (24-well inserts; pore size, 8µm; BD Bioscience) for invasion assays.
The medium in the top component was aspirated and replaced with serum-free media 2 h after seeding, and medium containing 20% serum was used as a chemotractant in the lower chambers. Cells were allowed to migrate for 24 h, and then the cells on both sides of the chamber were fixed with 50/50% acetone/methanol. Cells at the bottom of the chamber were stained with crystal violet (C3886, Sigma). Then, the picture of each well was taken to image the nuclei of the cells with the ×10 objective, and the cell nuclei in each field was counted using Image-J software (NIH; http://rsb.info.nih.gov/ij/).
Thank you very much for your kind response. This experiment has been designed using colon immortalized epithelial cell lines and CRC cells. My experimental cells are colon cancer cell-derived stem cells. Therefore, I think the number of the initial experiment is different rather than the conventional cell line, and the chemoattractant is also different.
There are straightforward 1 in all kits available these days with different kinds of ECM of choice. I use these regularly (from Sigma)
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