This core facility is committed to the large-scale cultivation of cell lines and preparation of different types of whole-cell, nuclear, and scaffold extracts as required for transcription factor identification, characterization and purification by each Pi of the program project. Different cell lines to be cultivated include: HeLa-S3, a nonlymphoid cell line adapted for growth in spinner cultures and commonly used for active transcription extracts and purification of ubiquitously-distributed transcription factors; JURKAT (TCR alpha/beta+) and PEER (TCR gamma/delta) T lymphoid cell lines for the analysis and isolation of TCF-2 and T cell lineage-restricted proteins that affect the silencing of the T cell receptor C-alpha enhancer; WEHI-23 B cells for the purification of rel-associated proteins, and smaller quantities of other cell lines, including PC-12 cells for CREB, and the Namalwa B cell line. Cells will be grown either in roller bottles (lymphoid cell lines) or spinner cultures (HeLa cells), and counted and split daily with fresh media to ensure logarithmic growth (essential for active transcription extracts). Laboratory space and some equipment has already been designated for this core, including a laminar flow hood and free-standing incubator, and, in addition there is a 37 degrees C warm room on our floor that holds several heavy-duty stirring plates for HeLa spinner cell cultures. Core D will be the overall responsibility of Dr. Jones, who will oversee the cell production schedule. The operation of the core will be supervised by Mr. Fernando Echeverria, who will count, split and harvest the cells, and establish the daily operating schedule of the facility. A second, part-time technical assistant, Mr. William Brown, will alternate the splitting and harvesting schedule with Mr. Echeverria, and will also be responsible for preparing and sterilizing the media, as well as ordering and stocking the tissue culture supplies. A part-time laboratory assistant is also requested to assist in daily autoclaving of the water for media, the bottles for media storage, as well as the large 6L flasks that are used to grow HeLa cells.
