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Unformatted text preview: Analysis of Cell Growth and Analysis of Cell Growth and Apoptosis Apoptosis Reference book #1: Chapter 6.3-6.4 Cell Cycle Cell Cycle Interphase : G1 phase S phase: DNA synthesis G2 phase M phase: DNA Replication Mitosis Cytokinesis The control of cell cycle: Two checkpoints The control of cell cycle: Two checkpoints Cell cycle is regulated by environmental cues Cell cycle is regulated by environmental cues Cell-cell interactions Microenvironment Chemical environment (e.g., growth factors, ECM, oxygen, nutrition) Mechanical environment (fluid, substrate) How many times can cells divide How many times can cells divide Embryonic stem cells (300-450 doublings) Cells in adult tissue (30-50 doublings) Quantify DNA content by using flow How to measure cell growth rate How to measure cell growth rate y y g cytometry Detect BrdU (bromodeoxyuridine) incorporation into DNA (in place of thymidine) Measurement of DNA content by flow cytometry Measurement of DNA content by flow cytometry etach cells G /G 1 Detach cells Stain cells in suspension with propidium iodide (PI) Quantify the fluorescence intensity of PI using flow S G 2 /M DNA Content cytometer 1. Incubate BrdU with cells for 1-2 hours (allowing BrdU being...
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- Fall '10
- Tissue Engineering