bis_104_sample_exam_key_07 - Page 2

bis_104_sample_exam_key_07 - Page 2 - Apply a fluorescently...

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PAGE 2 Now that you are confident that your cell cultures are healthy and free of bacterial contamination, you set about to test part (a) of your hypothesis. Design one in vitro experiment to test the idea that Schwann cells can induce undifferentiated endothelial cells to become arterial cells. Include a step by step procedure and describe the results you would anticipate if your hypothesis were correct. Describe one appropriate control for your experiment. EXPERIMENT: Obtain purified populations of Schwann cells and undifferentiated endothelial cells (PECAM-1 positive; EphrinB2 negative) by FACS 9we have not covered this. Mix the two cell types together in cell culture and let them interact for 2 -3 days.
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Unformatted text preview: Apply a fluorescently labeled antibody for arterial cell markers such as EphrinB2 or CX40. Wash away any unbound antibody and examine cells by fluorescence microscopy . RESULTS OF EXPERIMENT If differentiation has occurred, endothelial cells should show a positive reaction (fluorescence) with the anti-CX-40 and anti-EphrinB2 antibodies . CONTROL: Purify and culture PECAM-1 positive endothelial cells alone, without Schwann cells. Treat with labeled antibodies as in experiment and observe. RESULTS OF CONTROL No reaction (no fluorescense) with anti-arterial marker antibodies. Demo...
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