Gelatin-embedded cell-polymer constructs for histological cryosectioning.
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.
Brown
David A
DA
Department of Bioengineering, Henry L. Samueli School of Engineering, University of California, Los Angeles, California, USA.
Chou
Yu Fen
YF
Beygui
Ramin E
RE
Dunn
James C Y
JC
Wu
Benjamin M
BM
D000818:Animals; D002460:Cell Line; D016616:Cryoultramicrotomy / Q000295:instrumentation* / Q000379:methods*; D005780:Gelatin / Q000737:chemistry* / Q000494:pharmacology; D051379:Mice; D008856:Microscopy, Fluorescence; D011108:Polymers / Q000737:chemistry*; D000267:Tissue Adhesions; D023822:Tissue Engineering
D011108:Polymers; D005780:Gelatin