separating normal epithelial cells from malignant transformed - protocol suggestions? (Feb/09/2007 )
in an epithelium of MDCK we have some likely malignant transformants; they build clusters and grow in second an more layers; I like to separate those transfomed cells form the phenotype normal epithelium cells; as the transformed cells may have a loser cell-cell and cell-matrix contact, a differential splitting may help;
as we do not know any specific surface markers of the transformed cells, FACS sorting will not work at this early level of examination;
does anyone have a suggestion for a simple and effective protocol for separation or enrichement?
-The Bearer-
QUOTE (The Bearer @ Feb 9 2007, 02:39 AM)
in an epithelium of MDCK we have some likely malignant transformants; they build clusters and grow in second an more layers; I like to separate those transfomed cells form the phenotype normal epithelium cells; as the transformed cells may have a loser cell-cell and cell-matrix contact, a differential splitting may help;
as we do not know any specific surface markers of the transformed cells, FACS sorting will not work at this early level of examination;
does anyone have a suggestion for a simple and effective protocol for separation or enrichement?
as we do not know any specific surface markers of the transformed cells, FACS sorting will not work at this early level of examination;
does anyone have a suggestion for a simple and effective protocol for separation or enrichement?
Hello! I think may be this is one possible way not simple and may be delirious!
Try to take a chance and inoculate (10-12 million cells) into nude mice. If this cells take a road for malignacy may be they form tumors It is possible way to enrich your transformed population and then - isolation primary culture! 1 chance in 100
-circlepoint-
QUOTE (circlepoint @ Feb 13 2007, 09:03 PM)
QUOTE (The Bearer @ Feb 9 2007, 02:39 AM)
in an epithelium of MDCK we have some likely malignant transformants; they build clusters and grow in second an more layers; I like to separate those transfomed cells form the phenotype normal epithelium cells; as the transformed cells may have a loser cell-cell and cell-matrix contact, a differential splitting may help;
as we do not know any specific surface markers of the transformed cells, FACS sorting will not work at this early level of examination;
does anyone have a suggestion for a simple and effective protocol for separation or enrichement?
as we do not know any specific surface markers of the transformed cells, FACS sorting will not work at this early level of examination;
does anyone have a suggestion for a simple and effective protocol for separation or enrichement?
Hello! I think may be this is one possible way not simple and may be delirious!
Try to take a chance and inoculate (10-12 million cells) into nude mice. If this cells take a road for malignacy may be they form tumors It is possible way to enrich your transformed population and then - isolation primary culture! 1 chance in 100
thanks circlepoint, a nice idea; I think it will be more for our long range interest; I have to write an application for animal experiments
-The Bearer-