The primary purpose of this hands-on, laboratory-based course is to provide an understanding by students about how cultured cells can be easily manipulated in the laboratory and used to discover a vast array of molecular pathways governing cell growth and differentiation. Some of these exercises can also be adapted to studies aimed at testing different engineered modalities for cell and/or tissue biocompatibility or toxicity. Thus the skills learned by students will prepare them for a wide variety of academic or industrial careers that utilize cell culture technologies, ranging from testing basic cell growth, proliferation, or differentiation to designing and testing novel drug delivery platforms or cancer treatments.
OVERVIEW OF THE COURSE
INTRODUCTION TO CELL CULTURE
BRIEF REVIEW OF CELL CULTURE BIOLOGY
BASIC CONCEPTS FOR CELL CULTURE MAINTENANCE
EXERCISE #1: Basic Handling and Passage of Adherent Cells in Culture
EXERCISE #2: Influence of Serum, Coated Surfaces, and Alteration in Signal Transduction on Cell Division and Differentiation
EXERCISE #3: Immunocytochemistry
EXERCISE #4: Transient Transfection of Mammalian Cells
EXERCISE #5: Passage of Anchorage-Independent Cells in Suspension Cultures and Preparation for Cryopreservation
EXERCISE #6: Cryogenic Storage of Cells
EXERCISE #7: Snap Thaw of Anchorage-Independent Cells
EXERCISE #8: Differentiation of Human Leukemic HL-60 Cells
EXERCISE #9: Wright and Giemsa Staining of HL-60 Cells
EXERCISE #10: Immunolabeling of Cell Surface Markers and Staining of Nuclei for Assessment by Imaging Flow Cytometry
EXERCISE #11: Hematopoietic Stem Cell Isolation and Differentiation
EXERCISE #12: Embryonic Cortical Neuron Culture and Differentiation
EXERCISE #13: Independent Project
REFERENCES