This study was supported by: NIH R01HD29963 to D.D.C.; NIH U54HD007495 to S.M.H.; and NIH R01HD067721 to S.L.Y. and B.A.L. The authors have no competing interests to declare.

MUC4, a transmembrane glycoprotein, interferes with cell adhesion, and promotes EGFR signaling in cancer. Studies in rat models have demonstrated steroid hormonal regulation of endometrial MUC4 expression. In this study, qRT-PCR screening of mouse tissues determined that Muc4 mRNA also was robustly expressed in mouse uteri. Previous studies from our labs have demonstrated MUC4 mRNA was expressed at levels <1% of MUC1 mRNA in human endometrium and endometriotic tissue. Multiple human endometrial adenocarcinoma cell lines were assayed for MUC4 mRNA expression revealing extremely low basal expression in the Ishikawa, RL-95-2, AN3CA, and KLE lines. Moderate to high expression was observed in HEC50 and HEC-1A cells. MUC4 mRNA expression was not affected by progesterone and/or estrogen treatment, but was greatly stimulated at both mRNA and protein levels by proinflammatory cytokines (IFN-γ and TNF-α), particularly when used in combination. In endometrial tissue, MUC4 mRNA levels did not change significantly between normal or cancerous samples; although, a subset of patients with grade 1 and 2 tumors displayed substantially higher expression. Likewise, immunostaining of human endometrial adenocarcinoma tissues revealed little to no staining in many patients (low MUC4), but strong staining in some patients (high MUC4) independent of cancer grade. In cases where staining was observed, it was heterogeneous with some cells displaying robust MUC4 expression and others displaying little or no staining. Collectively, these observations demonstrate that while MUC4 is highly expressed in the mouse uterus, it is not a major mucin in normal human endometrium. Rather, MUC4 is a potential marker of endometrial adenocarcinoma in a subset of patients.

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.

We discuss and demonstrate why FTIR/ATR spectra can only be calibrated in wavelength, not intensity, for comparison with other data sets at present. This is because the intensity calibration must remove the instrument response function. To address this problem, we suggest a possible approach.