Research Project: iPSCs Differentiation into vSMCs

My research project for this summer is to model the protocol as outlined in the publication Generation of vascular endothelial and smooth muscle cells from human pluripotent stem cells (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4566857/) for differentiation of induced pluripotent stem cells (iPSCs) into vascular smooth muscle cells (vSMCs).

This is an overview of the differentiation process.

iPSCs are stem cells that are derived from adult cells. They grow in abundant numbers and grow more quickly than primary cells (cells collected as differentiated cells). This makes them ideal for use in disease and drug modeling. There are two pathways of differentiation using this protocol: synthetic and contractile. The synthetic condition results in more round, confluent cells that are not very responsive to external signals. The contractile condition–which is ideal for our purposes–results in cells that are more responsive to external stimuli, but do not grow as confluent as the synthetic cells.

My tasks are to replicate the protocol and then modify it, in order to optimize the amount of cells produced and the functionality of the cells. We do this by changing out different media formulations at different time points during the differentiation process. At the end of protocol, we check using immunostaining to see if certain proteins are being expressed by the cells, which allows us to see if the differentiation protocol has been successful.

This is an example of the Immunostaining that we have done so far.

There are various reasons for doing this project. For example, in optimizing the protocol the cells will be able to be cultured in greater volumes and be used for various disease models. These models could also be used to show the effects that certain drugs/medications have on the cells. The Truskey lab does a lot of disease/drug modeling, relating to the idea of microphysicological systems, so this type of project could also allow for more effective usage of lab resources.