Head and Neck Cancer

In this video, we describe the isolation of epithelial cells from a murine tongue. These cells can be further cultured to give rise to cell colonies.

This video demonstrates the steps involved in the pretreatment of FFPE tissue specimens for RNA chromogenic in situ hybridization. The pretreatment process helps maintain nucleic acid integrity within the cells and overall tissue morphology.

This video describes a sequential hybridization strategy that involves hybridizing mRNA probes to specific target mRNA. Following probe hybridization, signal amplification is performed to enhance resolution via reduction of signal-to-noise ratio during RNA-CISH of histological samples.

This video presents the protocol for signal detection during RNA chromogenic in situ hybridization. This technique can be used to diagnose an active oncogenic infection visually in an intact tissue specimen.

In this video, we demonstrate the extraction of dorsal root ganglion or DRG from a harvested rat spine for downstream applications.

This video describes the technique to embed 3D spheroids in agarose gel drops for preservation. The embedded 3D spheroids can be further used for downstream analysis.

This video demonstrates the preparation of the murine tongue extracellular matrix by using the decellularization method, which involves exposing the harvested tongue to a series of physical, chemical, and enzymatic treatments. The resulting TEM scaffold retains its native extracellular matrix composition, removing the cellular components.

In this video, we present the protocol to obtain finely ground extracellular matrix powder from lyophilized decellularized tissue by cryomilling. The powdered ECM retains its biochemical composition and can be used for microcarrier synthesis.

This video describes the preparation of a homogenized and digested extracellular matrix suspension from decellularized and lyophilized tissues. This technique helps in fabricating ECM-derived porous foams and microcarriers for use as substrates in advanced 3D cell culture models.

This video describes the method for synthesizing pure ECM-derived foams without the need for chemical crosslinking. These ECM-derived foams find applications as advanced 3D in vitro cell culture models or as pro-regenerative bioscaffolds.

This video demonstrates the procedure of extracellular matrix-derived microcarrier fabrication by electrospraying. These microcarriers simulate the native tissue-specific microenvironment and are stable in long-term 3D in vitro cell culture models.

In this video, we demonstrate the procedure for developing an in vitro perineural cancer cell invasion model by co-culturing mouse dorsal root ganglion and neurotropic cancer cells. The model can be utilized to conduct cancer assays, producing rapid and reproducible results.

In this video, we describe the isolation of a subpopulation of cancer stem-like cells from head and neck squamous carcinoma cell lines using fluorescence-activated cell sorter or FACS. The isolated side population of cells can be used for further downstream analysis.

In this video, we demonstrate an in vitro method for co-culturing DRG and cancer cells to study neurotropism and neural invasion.

This video describes the colorimetric assay, methyl thiazole tetrazolium (MTT) assay, to measure drug-induced cytotoxicity in cancer cells and determine their metabolic activity. This assay is based on the metabolic conversion of tetrazolium salts into colored products, reflecting the mitochondrial activity of cells.

This video describes the high throughput colorimetric screening assay, sulforhodamine B assay, to measure drug-induced cytotoxicity in cancer cells. The assay determines the drug resistance by measuring the cell density based on the cellular protein content.

This video demonstrates the methodology to screen for major cell death profiles in ionizing radiation (IR) induced cancer cell lines using DAPI staining assay. The DAPI-stained nuclei of target cells undergoing cell death exhibit distinct morphologies and can be easily identified by fluorescence microscopy.

In this video, we demonstrate the preparation of a decellularized pig esophagus scaffold, with its biochemical architecture and composition preserved, for tissue engineering applications.

This video describes an in vitro culture technique to generate human organotypic esophageal constructs by co-culturing primary human esophageal fibroblast and squamous epithelial cells within a decellularized porcine scaffold. These 3D culture models mimic the function and physiology of their in vivo tissue counterparts.

In this video, we demonstrate a procedure to isolate exosomes from target biofluids using magnetic microbeads. The procedure enables the rapid capture of exosomes, which can be used for downstream analysis.

This video presents the protocol for culturing salispheres, the three-dimensional spheroids derived from salivary glands. The salispheres can be further used to assess the impact of head and neck cancer on saliva production from salivary glands of affected human patients.

This video describes the use of low-magnification, dark-field microscopy to quantify specific exosomes in small volume biofluidic samples. The exosomes thus identified can serve as potential cancer biomarkers.

Source:Carly Fassler1, Liyu Huang1, Kavita Prasad1, Jiayi Xu2, Guansen Tong2, Jie Ying Wu2, Michael C. Topf1,31Department of Otolaryngology – Head and Neck Surgery, Vanderbilt University Medical Center.2Computer Science Department, Vanderbilt University.3Vanderbilt University School of Engineering, Vanderbilt University.This video demonstrates using augmented reality (AR) for tumor resection, where a 3D-scanned tumor specimen is visualized as a hologram on a head-mounted display. Surgeons can...

This video demonstrates the procedure of intravenously injecting cancer cells into the chicken chorioallantoic membrane vasculature to generate a CAM cancer metastasis model. The injected cells eventually form scattered single-cancer cell colonies, allowing for the study of cancer invasion and metastasis.

In this video, we describe the isolation of metastatic cancer cell colonies from the chicken chorioallantoic membrane (CAM) using shell-less egg culture. This method helps screen for factors that may be responsible for cancer metastasis.

In this video, we develop an orthotopic head and neck cancer murine model by intramucosal inoculation of the squamous cell carcinoma cell line. The tumor-bearing mouse would show resemblance to clinically relevant symptoms of human disease and can thus be used to study cancer progression and discover novel therapeutic strategies.

In this video, we describe the method to co-culture dorsal root ganglia and cancer cells on chick chorioallantoic membrane or CAM to study perineural invasion.

This video demonstrates the isolation of tumor-bearing chorioallantoic membrane from the chicken embryo. The harvested CAM can be used for further downstream applications.

This video demonstrates the in vivo delivery of miRNA inhibitors as therapeutic agents against tumor growth in orthotopic mouse models. The intratumoral administration of synthetic miRNA inhibitors or antagomiRs allows them to bind and inhibit endogenous miRNAs, thereby causing mRNA upregulation and suppression of tumor growth.

This video describes the procedure for orthotopic inoculation of thyroid cancer cells into the thyroid gland of mouse models, which may then serve as a valuable platform to assess carcinogenesis and discover potential treatment strategies.

In this video, we describe a step-by-step protocol to isolate spinal cord from a mouse model for in vitro studies.