title: Establishment of 3D Co-Culture Models from Different Stages of Human Tongue Tumorigenesis: Utility in Understanding Neoplastic Progression.
abstract: To study multistep tumorigenesis process, there is a need of in-vitro 3D model simulating in-vivo tissue. Present study aimed to reconstitute in-vitro tissue models comprising various stages of neoplastic progression of tongue tumorigenesis and to evaluate the utility of these models to investigate the role of stromal fibroblasts in maintenance of desmosomal anchoring junctions using transmission electron microscopy. We reconstituted in-vitro models representing normal, dysplastic, and malignant tissues by seeding primary keratinocytes on either fibroblast embedded in collagen matrix or plain collagen matrix in growth factor-free medium. The findings of histomorphometry, immunohistochemistry, and electron microscopy analyses of the three types of 3D cultures showed that the stratified growth, cell proliferation, and differentiation were comparable between co-cultures and their respective native tissues; however, they largely differed in cultures grown without fibroblasts. The immunostaining intensity of proteins, viz., desmoplakin, desmoglein, and plakoglobin, was reduced as the disease stage increased in all co-cultures as observed in respective native tissues. Desmosome-like structures were identified using immunogold labeling in these cultures. Moreover, electron microscopic observations revealed that the desmosome number and their length were significantly reduced and intercellular spaces were increased in cultures grown without fibroblasts when compared with their co-culture counterparts. Our results showed that the major steps of tongue tumorigenesis can be reproduced in-vitro. Stromal fibroblasts play a role in regulation of epithelial thickness, cell proliferation, differentiation, and maintenance of desmosomalanchoring junctions in in-vitro grown tissues. The reconstituted co-culture models could help to answer various biological questions especially related to tongue tumorigenesis.
authors: Vaidya Laboratory, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, Maharashtra, India.
authors: Sharada
authors: S
authors: Sawant
authors: Vaidya Laboratory, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, Maharashtra, India.
authors: Harsh
authors: H
authors: Dongre
authors: Vaidya Laboratory, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, Maharashtra, India.
authors: Archana Kumari
authors: AK
authors: Singh
authors: Vaidya Laboratory, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, Maharashtra, India.
authors: Shriya
authors: S
authors: Joshi
authors: Gade Laboratory for Pathology, Department of Clinical Medicine, and Center for International Health, Department of Global Public Health and Primary Care, University of Bergen, Bergen, Norway.|Department of Pathology, Haukeland University Hospital, Bergen, Norway.
authors: Daniela Elena
authors: DE
authors: Costea
authors: Vaidya Laboratory, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, Maharashtra, India.
authors: Snehal
authors: S
authors: Mahadik
authors: Vaidya Laboratory, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, Maharashtra, India.
authors: Chetan
authors: C
authors: Ahire
authors: Nups and Sumo Biology Group, Department of Biological Sciences, Indian Institute of Science, Education and Research, Bhopal, Madhya Pradesh, India.
authors: Vidhi
authors: V
authors: Makani
authors: Vaidya Laboratory, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, Maharashtra, India.
authors: Prerana
authors: P
authors: Dange
authors: Oral Surgery, Head and Neck Unit, Tata Memorial Hospital (TMH), Parel, Mumbai, India.
authors: Shilpi
authors: S
authors: Sharma
authors: Oral Surgery, Head and Neck Unit, Tata Memorial Hospital (TMH), Parel, Mumbai, India.
authors: Devendra
authors: D
authors: Chaukar
authors: Vaidya Laboratory, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, Maharashtra, India.
authors: Milind
authors: M
authors: Vaidya
mesh_terms: D002454:Cell Differentiation; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic; D018920:Coculture Techniques / Q000379:methods*; D051182:Desmogleins / Q000378:metabolism; D051180:Desmoplakins / Q000378:metabolism; D003896:Desmosomes / Q000378:metabolism / Q000648:ultrastructure; D005347:Fibroblasts / Q000473:pathology; D006801:Humans; D015603:Keratinocytes / Q000473:pathology; D014059:Tongue / Q000473:pathology; D014062:Tongue Neoplasms / Q000473:pathology*; D051185:gamma Catenin
chemical_list: C495294:DSP protein, human; D051182:Desmogleins; D051180:Desmoplakins; C495292:JUP protein, human; D051185:gamma Catenin