[Separation of adhering cell colonies with a direct digestion method].
OBJECTIVE
To establish a highly efficient method for the separation of adhering cell colonies.


METHODS
Cell suspension was diluted gradiently to four different densities and then seeded on a cell culture plate covered with parted fibrinous membranes. After colonies appeared, they were digested directly with 0.25% trypsin and picked out with a pipette tip.


RESULTS
At the density of 5-10.cm-2, colonies could be separated most efficiently in both continuous and normal diploid cell line colonies. The cell number could reach 10(6) after one month of culture for a continuous cell line colony and one and a half month of culture for a normal diploid cell line colony.


CONCLUSIONS
With the direct digestion method, single cell colonies can be effectively separated from the plate covered with parted fibrinous membranes, when the cell was initially seeded at the density of 5-10.cm2.
Zhao
Di-cheng
DC
National Laboratory of Medical Genetics, Central South University, Changsha 410078, China.
Long
Zhi-gao
ZG
Zheng
Duo
D
D000818:Animals; D016466:CHO Cells / Q000166:cytology*; D002469:Cell Separation; D002478:Cells, Cultured; D006224:Cricetinae; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D013234:Stem Cells / Q000166:cytology*; D014471:Umbilical Veins / Q000166:cytology