Gelatin-embedded cell-polymer constructs for histological cryosectioning

Journal of Biomedical Materials Research. Part B, Applied Biomaterials
David A BrownBenjamin M Wu

Abstract

Many tissue-engineering strategies involve the delivery of cells via porous polymer scaffolds. Obtaining histological sections of the emerging tissue is often necessary to analyze numerous characteristics of the microscopic environment. However, difficulties arise upon applying standard histological techniques to cell-seeded polymer scaffolds. This report describes a simple and reliable method for cryosectioning cell-polymer constructs embedded in gelatin. Solvent-soluble (PLGA) and insoluble (PGA) scaffolds were cultured in vitro with preosteoblasts, followed by histological processing with paraffin, OCT, or gelatin. Although paraffin-embedded PGA scaffolds withstood standard sectioning and rinsing steps, paraffin-embedded PLGA scaffolds were partially dissolved during the clearing step. OCT-embedded scaffolds produced sections that did not adhere well to slides, and most of the sample was lost during rinsing steps. In contrast, gelatin-embedded scaffolds exhibited adequate structural integrity during cryosectioning, adhered well to the slides, retained the actual polymer morphology, and exhibited compatibility with common stains.

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Citations

May 6, 2006·Tissue Engineering·James C Y DunnBenjamin M Wu
May 9, 2008·Regenerative Medicine·James Cy Dunn
Feb 29, 2008·Biotechnology and Bioengineering·J V NygaardF Besenbacher
Feb 11, 2011·Biotechnology and Bioengineering·Igor KuzinAndrea Bottaro
Dec 4, 2009·Journal of Biomedical Materials Research. Part B, Applied Biomaterials·Edward A PhamBenjamin M Wu
Jun 7, 2008·Clinical Orthopaedics and Related Research·Eugene FarngDavid R McAllister
Jul 8, 2005·Journal of Biomedical Materials Research. Part B, Applied Biomaterials·Yu-Fen ChouBenjamin M Wu
Dec 19, 2018·Histochemistry and Cell Biology·Julia FuchsDagmar Brislinger

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