I came across HT-29 from different laboratories. When I grow them, one gave me epithelial-like shaped while the other one look more rounded and will grow continuously into sheet-like layer of cells. I am confused which is the right ones. Both cell line have passage number of less than 10. Would appreciate any help. Thanks.
Originally they look different at low and high densities and I think crossing the critically high densities several times and repeated splitting could lead to stable morphology changes. I also have lines which have different morphologies. Both of them are epithelial, but clearly differ. There was also visible difference in their adherence properties. We will have to consider the ATCC picture as the standard appearance:
http://www.atcc.org/products/all/HTB-38.aspx#characteristics
Originally they look different at low and high densities and I think crossing the critically high densities several times and repeated splitting could lead to stable morphology changes. I also have lines which have different morphologies. Both of them are epithelial, but clearly differ. There was also visible difference in their adherence properties. We will have to consider the ATCC picture as the standard appearance:
http://www.atcc.org/products/all/HTB-38.aspx#characteristics
Hi Richard,
I agree with Manoj. I have worked with another colorectal cancer cell line, the Caco-2, and they also look different at high densities. They look like round shapped initially but later their appearance is like the image I also enclose:
http://www.lgcstandards-atcc.org/~/media/EF14D782C6DC4D6389314E377EF593A3.ashx
There is no "standard appearance". It depends on time in culture, cell density, the medium you grow the cells in and experimental factors. See Cell Differ Dev. 1990 Sep;31(3):207-18. They tend to form islands of cells when subconfluent and tightly polarized monolayers when confluent for some time. If you want to study them as functional epithelial cells you need to let them polarize. Macroscopically you can see that this has happened when the confluent monolayer starts to "dome", i.e. produce large round patches of cells that are bulging away from the plastic surface (due to transepithelial fluid secretion from the medium to the plastic surface, under an impermeable monolayer). If you grow them on permeable transwell supports you can measure a large increase in transepithelial resistance when polarization has occurred (which requires both full confluency and tight junction formation). If you have not grown polarized epithelial cells before, this seems counterintuitive, as with most other cell lines you want to avoid confluency, but if you want to study polarized epithelial cells it is a prerequisite. During the confluent polarization period you need to change the medium more frequently. Another important point: Whenever you get unverified cell lines from other laboratories you must fingerprint them, so that you know they are actually what you think they are. Even ATCC has had a long history of cross-contaminated cell lines. You will save yourself a lot of wasted time, and also some journals now are requiring fingerprinting of cell lines before they will accept papers where conclusions rely heavily on observations in cultured cells.
Thank you Manoj, Elena and Norbert. Your suggestions are of great help to me. I am culturing CACO-2 as well and they also gave me morphology as described by Elena. My research team do not have a tradition of doing cell line fingerprinting, but I agreed that cross contamination of cell line do easily happen, especially from poor hand writing.
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