title: Validation of a postfixation tissue storage and transport medium to preserve histopathology and molecular pathology analyses (total and phosphoactivated proteins, and FISH).
abstract: Tumor biomarker studies are integral to oncology clinical trials but may yield artifactual results owing to variation in sample procurement and processing. Ethanol, 70% vol/vol, was validated as a sample transport medium using markers of the PI3K/Akt/mTOR pathway. BT474 tumor xenografts were excised and slices were immediately placed into formaldehyde and fixed for 24 hours. Fixed tissue slices were immediately processed into paraffin or transferred to 70% vol/vol ethanol and stored at room temperature for 1, 2, and 4 weeks before further processing. Freshly cut tissue sections were evaluated for pAKT(S473), HER2, pHER-2(Y1248), pS6(S235/236), and pS6(S240/244), Ki-67, and HER2 by fluorescence in situ hybridization and stained with H&E and Masson trichrome. No significant changes were observed when comparing samples stored in 70% ethanol for up to 4 weeks with immediately processed tissue. Ethanol, 70% vol/vol, provides a safe storage medium for formaldehyde-fixed tumor tissue, facilitating sample transport during multicenter clinical trials.
authors.affiliation: Oncology Research, Novartis Institutes of Biomedical Research, Novartis Pharma AG, Basel, Switzerland.
authors.forename: Michael M
authors.initials: MM
authors.lastname: Stumm
authors.forename: Maja R
authors.initials: MR
authors.lastname: Walker
authors.forename: Caroline
authors.initials: C
authors.lastname: Stork
authors.forename: Noelle
authors.initials: N
authors.lastname: Hanoteau
authors.forename: Urs
authors.initials: U
authors.lastname: Wagner
authors.forename: Terence M
authors.initials: TM
authors.lastname: O'Reilly
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism; D045744:Cell Line, Tumor; D004247:DNA / Q000032:analysis / Q000737:chemistry; D000431:Ethanol / Q000737:chemistry; D005260:Female; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D016591:Histocytological Preparation Techniques / Q000379:methods*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008325:Mammary Neoplasms, Experimental / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009363:Neoplasm Proteins / Q000378:metabolism; D057089:Pathology, Molecular / Q000379:methods*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D013048:Specimen Handling / Q000379:methods*; D013997:Time Factors
chemical_list: D014408:Biomarkers, Tumor; D004247:DNA; D000431:Ethanol; D005404:Fixatives; D005557:Formaldehyde; D009363:Neoplasm Proteins; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human