AfCS > AfCS laboratories > Cell Preparation and Analysis Laboratory

ALLIANCE FOR CELLULAR SIGNALING

CELL PREPARATION AND ANALYSIS LABORATORY

University of Texas Southwestern Medical Center
Dallas, TX

Overview

The general mission of the Alliance for Cellular Signaling (AfCS) Cell Preparation and Analysis Laboratory is to produce consistent and timely preparations of cells for experimental use by AfCS laboratories and to perform experiments that have been designed by AfCS investigators.

There are three major areas of focus. A top priority for the first years of the AfCS effort was the establishment of methods and capacities for large-scale preparations of mouse splenic B cells and the culture of selected mouse cell lines. Current effort is directed solely on the culture of RAW264.7 cells. All preparations of cells must be reproducible both within and between laboratories, meet defined specifications, and be carried out to meet demand on a scheduled basis.

The Cell Laboratory is responsible for a variety of cell-based assays, chosen to measure the response of cells to extracellular stimuli. Samples produced by the laboratory are either analyzed within the lab or sent to other AfCS laboratories for more detailed analysis. In the initial years of study, the focus was on the ligand/input screen to determine the complexity of cellular responses to extracellular ligands. In the last year, focus is shifting to the use of similar or more targeted assays that seek to define intracellular signaling networks by perturbation analysis.

A third priority is the establishment of new methodology to assess information flow through other molecular nodes not yet targeted by AfCS assays. Emphasis for new targets is developed by the Systems committee and modeling efforts.

In order to accomplish these objectives, the laboratory is organized in teams centered on planning, production, and development. Goals for personnel include the development of expertise in multiple areas, leadership in experimental implementation, high standards of quality, and close coordination with other AfCS laboratories.

Staff

Alliance for Cellular Signaling
Cell Preparation and Analysis Laboratory
University of Texas Southwestern Medical Center
5323 Harry Hines Blvd., J5-126
Dallas, Texas 75390-9196
Phone: 214-648-2271
Fax: 214-648-5006

Director	Paul Sternweis, Ph.D.
Research Scientist	Dianne DeCamp, Ph.D.
Research Scientist	Robert Hsueh, Ph.D.
Research Scientist	Lily Jiang
Research Scientist	Keng-Mean Lin, Ph.D.
Research Assistant	Julie Collins
Research Assistant	Richard Davis
Research Assistant	Audra Wendt
Research Assistant	Amy Pope
Research Assistant	Blake Pond
Research Assistant	Angela Carter
Research Assistant	Natalie Strickland
Research Assistant	Elaine Campbell
Lab Administrator	Kimberly Edwards
Data Manager	Lonnie Sorrells
Lab Aid	Philip Ramirez

Slide Presentations from May 23-26, 2004, AfCS Annual Meeting

• "Calcium and Cyclic AMP", Paul Sternweis
• "Phosphoproteins/Intermediate endpoints", Lily Jiang

Goals for Year 5 (9/04-8/05)

• Complete the dual ligand screen in RAW264.7 cells with phosphoproteins as the output.
• Design and perform limited dual ligand treatments of RAW264.7 cells for transcript and lipid analysis as requested.
• Cooperate in the design and performance of specific experiments in support of modeling efforts.
• Treat control and knock-down cells with designated ligands and prepare samples for analysis of desired outputs. Perform assays on such samples when appropriate.
• Initiate or cooperate in the development of new output assays for assessment of perturbation paradigms and modeling efforts.