Question

It is desired to identify all possible effects of a particular drug on selected adult stem cell pools. All kinds of equipment and molecular biology tools can be used for this project and all the required experiments. Please write down all the steps to reach that goal.

Answer 1

To study effect of any drug on adult stem cell . We can study it in following point

1, morphological study- here we can study the effects of drug on adult stem cell morphology ( change in cell structure) by using microscope.

We can simply do H and E stain to study the morphological and histological changes .

2. Effect of durg on adult stem cell proliferation

This we can study by simple count ate cell no at different time interval of durg concentration. We can also do MTT or WST assay , PI staining for this.

3. Proteomics studies- we can study alteration in gene expression in response to a drug in adults stem cell. By doing proteomic studies.

4, this can be further validated by examining expression of mRNA using QPCr ( quantitative PCR), whose protein showed alteration in the proteomic study.

5, we can study effect of drug on various enzyme by measuring their enzymatic activity.

6, if this drug is affecting gene expression, one reason might be its affecting methylation status. This can be studied using Methylation specific PCR.