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Stem Cell Projects

A video produced by our lab for the EFRI conference

This video introduces some of the key stem cell related projects in our group including our work with linking cardiac differentiation with physical and chemical cues from substrates and soluble factors and our work investigating Wnt signaling in hPSCs confined in microwells. This video was produced as part of the NSF initiative for emerging frontiers in research and innovation EFRI.(Courtesy of Chet Hsiao, Xiaojun Lian & Laurie Hazeltine)

Epithelial Differentiation and Skin Tissue Engineering

Cornified Cells

Our group has developed a protocol for differentiating epithelial cells from human pluripotent stem cells (hPSCs) using retinoic acid. We are developing methods to develop skin tissue in vitro from these epithelial cells. We are also investigating other epithelial cell types using our differentiation protocol.

The picture shows a H&E stain of hPSC derived keratinocytes seeded on primary fibroblasts and subjected to cornification in vitro . The formation of cornified cells at the upper surface indicates the formation of skin-like tissue.(Courtesy of Josh Selekman)

Wnt Signaling in Microwell Confinement

Our lab is investigating the effects on Wnt signaling when hPSCs are confined to microwells. We fabricate microwells, small cuboidal chambers for growing cells which have a side length of about 100 - 300 μm. These are fabricated on polymers and then the sides are coated with gold while the bottom is functionalized with extracellular matrix proteins. Our group has noticed that cells differentiate differently from cells grown in 2D cultures.

Cardiac Differentiation

Force map of cardiomyocyte

We are currently investigating the effects of some chemical and physical cues on cardiac differentiation from hPSCs. We are investigating the effects of various soluble factors, several ECM proteins and substrate stiffness on cardiogenesis. We have developed a technique for quantifying the beating forces in individual beating cardiac cells using hydrogels.

The video shows a beating cardiomyocyte differentiated from hPSCs, along with the green fluorescent beads embedded in the gel below. The movements of the beads can be quantified to calculate beating forces. This calculation leads to the force map shown alongside. (Cardiomyocytes differentiated by Xiaojun Lian, microscopy and image analysis by Laurie Hazeltine)