Question

4. (15) You designed a polymer based hydrogel scaffold which can direct neural stem cells (NSCs) to differentiate into neurons in a linear grid. To check whether the cells differentiated, you used immunofluorescence to stain the cells for neural markers MAP-2 and DCX and NSC marker Sox2. a) What data do you expect to see from your immunofluorescence experiment if the NSCs had differentiated? b) Is this experiment sufficient? What other experiments should we do to confirm? (No need to state specific experimental procedures, rather please focus on the biologically relevant information we wish to gather. Hint > what did the papers we read in class do?) 5. (10) You patterned ONE cell in EACH of the following shapes, assuming the cell volume is slightly greater than the volume of your pattern; please draw what you expect the actin network to look like. Circle where you think the cell would form large focal adhesions

Answer 1

4. (a) During the differentiation of neural stem cell, MAP-2 maker present in the full mature shows green fluorescent colour and DCX shows a dark brown colour. Differentiated cells are elongated cell. Sox indicates stem cell shows a stained circular differentiated cell (self-renewal).

(b) Yes, this experiment is sufficient. MAP-2 (microtubule-associated proteins) are the protein associated with microtubules during cell division to provide stable network structure. They are used as a marker for neural cells (perikarya and dendrites). DCX (doublin or lissencephalin-X) it is neuronal migration protein doublecortin, a microtubule-associated protein synthesized by neuronal stem cells and immature neurons. Sox2 is a protein molecule act as a transcription factor for the regulation and maintenance of stem cell.

5. small size cells are stem and immature neural cells. Mature cells are elongated nerve cells. Figure attached cell.

MAP Sox