Reassembly of Functional Human Stem/Progenitor Cells in 3D Culture

This chapter focuses on the culture of primary human cells from the salivary glands, typically parotid but also submandibular, where specialized acinar cells produce most of the components found in saliva and the intercalated ducts followed by striated ducts transport saliva to the oral cavity. Compared to many other epithelial cells, the zymogen-filled salivary acinar cells are very fragile, hence specialized techniques are needed to isolate and culture them. To reestablish the function of implantable 3D reassembled glands using tissue engineering approaches, it is critical to culture these cells in human-based matrices that permit them to move, reassemble, interconnect, and establish proper polarity by producing a basement membrane. Our team is working to develop a biologically based, implantable salivary gland replacement tissue for head and neck cancer patients suffering from post-radiation xerostomia using a "bottom up" reassembly paradigm. We use specialized extracellular matrix and growth factor supplemented hyaluronate hydrogels to promote reassembly of human salivary stem/progenitor cells (hS/PCs) isolated after surgical resection, a method we describe in this chapter. Cell-specific biomarkers are used to track the formation of the three major epithelial cell types comprising the salivary gland: acinar, ductal, and myoepithelial.