ABSTRACT
We used laser microdissection and pressure catapulting (LMPC) to isolate and subsequently recultivate cells from the colorectal cancer cell line HCT116. Cell isolation and recultivation was repeated 5 times from the preceding cell culture. The chromosomal composition of the HCT116 cells was analyzed at the beginning and after each recultivation. We did not observe any additional chromosomal structural or copy number changes during this procedure. Our data suggest that the procedure of cell isolation by LMPC and subsequent recultivation does not affect the karyotype of the respective cells. This technology will facilitate the analysis of different cell populations in a heterogeneous tumor cell population.
Subject(s)
Cell Culture Techniques/methods , Colorectal Neoplasms/genetics , Cell Separation , Colorectal Neoplasms/pathology , HCT116 Cells , Humans , Karyotyping , Microdissection , Nucleic Acid HybridizationABSTRACT
Laser micromanipulation systems are used worldwide in the field of life science research. Most of their applications focus on the isolation of specific cells from different types of tissue and the manipulation of subcellular structures within fixed or living cells. Using the PALM MicroBeam, it is possible to microdissect living cells from a cell culture, to catapult them into collection devices, and to re-cultivate the isolated cells. For this purpose, new protocols and special equipment were developed. It has also been demonstrated that Laser Microdissection and Pressure Catapulting (LMPC) have no influence on the proliferation rate of the cells. Even re-cultivated cell colonies, trypsinized and seeded out again, are still viable after a second LMPC-procedure. This new approach opens a wide field of interesting applications in cell biology, molecular pathology, and pharmacology.
