Histological Evaluation of Long-Term Collagen Type I Culture

Marcella Massimini; Mariarita Romanucci; Raffaella De Maria; Leonardo Della Salda

doi:10.1007/978-1-0716-2403-6_10

Histological Evaluation of Long-Term Collagen Type I Culture

Methods Mol Biol. 2022:2514:95-105. doi: 10.1007/978-1-0716-2403-6_10.

Authors

Marcella Massimini¹, Mariarita Romanucci², Raffaella De Maria³, Leonardo Della Salda²

Affiliations

¹ Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy. mmassimini@unite.it.
² Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy.
³ Department of Veterinary Sciences, University of Turin, Turin, Italy.

PMID: 35771422
DOI: 10.1007/978-1-0716-2403-6_10

Abstract

Histological approach to long-term culture on collagen type I permits the evaluation of vasculogenic mimicry morphological features and the identification of endothelial-like cell-specific antigens. Here, we show the preparation of collagen type I solution, the embedding and the sections cutting of D17 osteosarcoma cells long-term culture, and then the hematoxylin and eosin (H&E) staining to identify endothelial-like structure. Moreover, we provide the protocols for periodic acid-Schiff (PAS) staining to evidence glycoproteins and CD31 immunohistochemistry to exclude the presence of this endothelial marker, as per definition by vasculogenic mimicry concept. This method allows to consider long-term culture as tissue, promoting the deeper study of vascular-like structures in vitro.

Keywords: Collagen type I; In vitro markers; Paraffin-embedded long-term culture; Vasculogenic mimicry.

MeSH terms

Bone Neoplasms*
Collagen Type I
Humans
Immunohistochemistry
Neovascularization, Pathologic / pathology
Osteosarcoma*

Substances

Collagen Type I