CORE--FLUORESCENCE ACTIVATED CELL SORTING

$0P01FY2001CANIH

University Of Texas Md Anderson Can Ctr, Houston TX

Investigators

Linked publications & trials

Paper 38856690 Paper 37725573 Paper 37038957 Paper 33438761 Paper 31127149 Paper 30674535 Paper 30337364 Paper 29655803 Paper 29490944 Paper 29463558 Paper 29435107 Paper 29232553 Paper 29137349 Paper 28659338 Paper 28388555 Paper 28270436 Paper 27391151 Paper 27268264 Paper 27129202 Paper 26956049 Paper 26838606 Paper 26822154 Paper 26752456 Paper 26704388 Paper 26674329 Paper 26431162 Paper 26340096 Paper 26183077 Paper 26095711 Paper 26045609 Paper 26031918 Paper 25867648 Paper 25598437 Paper 25558171 Paper 25530214 Paper 25406351 Paper 25223583 Paper 25079338 Paper 24858343 Paper 24766216 Paper 24739393 Paper 24737502 Paper 24727332 Paper 24628293 Paper 24619500 Paper 24619497 Paper 24599548 Paper 24596420 Paper 24526787 Paper 24510534 Paper 24346116 Paper 24337870 Paper 24170028 Paper 24154679 Paper 24144307 Paper 24092807 Paper 23841653 Paper 23838935 Paper 23691988 Paper 23676502 Paper 23669290 Paper 23611563 Paper 23565503 Paper 23564911 Paper 23530930 Paper 23397500 Paper 23345254 Paper 23333150 Paper 23266036 Paper 23260083 Paper 23247046 Paper 23036488 Paper 23005582 Paper 22893484 Paper 22892554 Paper 22826565 Paper 22826139 Paper 22796535 Paper 22785211 Paper 22687754 Paper 22645176 Paper 22585577 Paper 22498477 Paper 22490334 Paper 22368688 Paper 22363446 Paper 22249254 Paper 22136251 Paper 22100978 Paper 22064351 Paper 22033489 Paper 22023523 Paper 22005650 Paper 21988645 Paper 21986948 Paper 21949692 Paper 21904380 Paper 21868571 Paper 21853076 Paper 21767516

Abstract

The FACS and FISH Core provides phenotypic cell cycle and genotypic analysis and separated cell populations to the investigators of this program project. The laboratory has developed cutting edge techniques in single cell analysis. For the quantitation of apoptotic cells, the TdT-b- dUTP assay (TUNEL) was modified for multi-parameter analysis in combination with DNA and BudR measurements (cell cycle, ploidy). Also, immunophenotypic analysis of dendritic cells (Project 4) and of progenitor cells, analysis of intracellular proteins related to apoptosis (bcl-2, bax) (Project 1), and of cell surface antigens including fas and MDR1 is provided. Expression of new regulators of apoptosis (Project 2) will be analyzed in normal and leukemic cells. Binding of Annexin V to phosphatidyl serine (PS) is utilized for testing changes in the membrane lipid structure associated with apoptosis. New assays to measure changes in the mitochondrial membrane potential and of cytochrome c in cells initiating apoptosis (CMXRos) and the detection of cleaved caspase 3 (Caspa-Tag Ph-phi Lux) are conducted for Projects 1, 4 and Core A. Quantitation of cellular antigens allows us to determine the Antibody Binding Capacity (ABC). Cd34 cells are MACS separated for subsequent analysis by FACS. Cell kinetic changes are determined in vitro by DNA/Ki67/CD34/CD38 FCM and the number of actual cell divisions is determined by pkh26 labeling., with cells undergoing one or up to ten divisions being separated by FACS for subsequent molecular analysis (Project 1). Cell cycle kinetics is determined in patients by infusions of IudR and BudR (Projects 1 and 3, Core A). Fluorescence in situ hybridization (FISH) determines the number of clonal leukemic interphase or metaphase cells. The combination of FISH and TUNEL of PS/Annexin V assay allows to discriminate apoptosis in normal and leukemic cells (Projects 1 and 4). Finally, progenitor and stem cell compartments (CD34+38-, CD34+KDR+ and CD34-lin- cells that eliminate Hoechst 33342, so-called "SP" cells) are sorted for investigations of XIAP and survivin in Project 1 and are analyzed for the presence of clonal leukemic and normal cells. The core provides critical support for all Projects and for Core A (clinical trials).

View original record on NIH RePORTER →