NCI Pediatric Preclinical Testing Program (PPTP) In Vitro Laboratories

The NCI Pediatric Preclinical Testing Program (PPTP)
Cell Culture Laboratory

Director:    Min H. Kang, PharmD
Co-investigator:    C Patrick Reynolds, MD PhD

The PPTP In Vitro laboratory is located at the Texas Tech University Health Sciences Center School of Medicine Cancer Center and is the cell culture (in vitro) portion of the NCI PPTP. The lab tests drugs on a panel of cell lines includes neuroblastomas, rhabomyosarcomas, lymphomas, ALL, AML, ESFT, and brain tumor cell lines).

PPTP In vitro Laboratory

Lab Layout Dr. Kang's laboratory conducts the in vitro preclinical testing for the PPTP. The assay employed is DIMSCAN, a fluorescence-based multi-well assay with a dynamic range of 4 logs of cell kill. The in vitro testing lab is equipped with 3 biological safety cabinets, a robot pipettor, 4 incubators (O2 control capable), a DIMSCAN, centrifuges, -80° and 4° GLP storage, and access to the COG Cell Culture Repository.

Methods employed:

The primary cytotoxicity assay for the lab is the DIMSCAN 96 well assay. DIMSCAN employs fluorescein diacetate (FDA) to identify viable cells, with quenching of fluorescence by eosin Y, and viable cells quantified using a DIMSCAN digital image microscopy system. (1,2). The DIMSCAN assay has a 4 log dynamic range when testing solid tumor (3, 4) and leukemia (5) cell lines. All cell lines used are human, have been tested and shown to be free of mycoplasma, and have been demonstrated to be unique using short tandem repeats (8).

Laboratory Funding

NCI PPTP Grant