Stable Cell lines

Sailesh Surapureddi Saisu at mcb.liu.se
Tue Nov 10 11:24:48 EST 1998



My Dear Expressionists

	I am trying to make some stable cell lines. I have tranfected my
constructs (has amp and neo as markers) to fibroblasts (Ca-Phos method).
After the removal of calcium precipitate, cells were split and seeded in
low density and treated with Neomycin sultfate (500 microgram/ml). The
control cells after sometime die out. But intrestingly all the lines which
I am trying to develop have confluent cells but the intresting thing is
that there are lot of
"islets of cells" with clear demarcations (they look kinda dark in color
when compared to the other cells in normal microscope observations.)

My Q is there an kind soul who knows what I am talking about or observed
such islets when making stable cell line ?

2. Can anyone suggest me on how to pick up individual islets without
disturbing the rest (I can only see them under microscope and not
distinguiable to normal vision)???


All your help is greatly appreciated in advance


Sailesh.



Sailesh Surapureddi, Ph.D
				Communicators
Plan #12, Cellbiologi		Work	+46-13-223917
Dept. of Biomedicine & Surgery  Res	+46-13-274718
Linköpings University		Cel	+46-70-4916570
S 581 85, Linkoping		Fax	+46-13-224149
Sweden.				Email  Saisu at mcb.liu.se





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