Lecture_3-_Deriving_neurons_from_ESCs

Lecture_3-_Deriving_neurons_from_ESCs - Phase contrast...

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If you look at your syllabus, you will notice that we had our Q/A with Dr. Sun scheduled for Thursday, April 28. We will change the Q/A to Tuesday, May 3 and will have the Minor Info Session on April 28 instead. Note The Following Change Still due on May 3 rd .
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Tri-lineage differentiation from human ESC-derived NPCs So what exactly are we looking at? Immunofluorescent staining for differentiation markers.
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Step 1 differentiation from human ESC to NPCs Nestin: labeled in red, protein found in cytoplasm. Sox2 labeled in green, protein found in nucleus. Hoechst blue, stains DNA (nucleus). hESC NPC terminal differentiated state (neuron or glia)
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Step 2 differentiation from human NPCs to neurons MAP2: labeled in green, protein found in cytoplasm. Ki67 labeled in red, protein found in nucleus. [Note: marker for cell division, not for neurons] Hoechst blue, stains DNA (nucleus). hESC NPC terminal differentiated state (neuron or glia)
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Unformatted text preview: Phase contrast (light microscopy), showing the morphology of the differentiated neurons. When passing white light through a biological specimen, the image is often hard to see: Phase Contrast But by manipulating how the light enters/exits the specimen, the contrast of the image can be enhanced. Step 2 differentiation from human NPCs to glia GFAP: labeled in green, protein found in cytoplasm. DAPI blue, stains DNA (nucleus). hESC NPC terminal differentiated state (neuron or glia) O4: labeled in green, protein found in cytoplasm. CNPase: labeled in red, protein found in cytoplasm. DAPI blue, stains DNA (nucleus). hESC hESC-NPC hESC-neuron Purification of human neurons from human ESCs Light microscopy (phase contrast) of ESCs differentiating. Differentiating ESCs entails changing the environmental signals...
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Lecture_3-_Deriving_neurons_from_ESCs - Phase contrast...

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