Specimen quality evaluation in Canadian biobanks participating in the COEUR repository.
Human biological specimens are important for translational research programs such as the Canadian Ovarian Experimental Unified Resource (COEUR) funded by the Terry Fox Research Institute. Sample quality is an important consideration, as it directly impacts the quality of ensuing research. The aim of the present study was to determine the quality of tissues collected from different sites contributing to the COEUR cohort. Samples from high-grade serous ovarian tumors (fresh frozen and corresponding paraffin-embedded tissues) were provided by nine participating Canadian biobanks. All samples were shipped to a central site using a Standard Operating Protocol (SOP). DNA and RNA extraction was conducted by the quality control division of the Canadian Tumor Repository Network (CTRNet). DNA quality was determined by ß-globin gene PCR amplification, and RNA quality by the RNA integrity number (RIN), as measured by the Agilent BioAnalyzer. DNA of acceptable quality had at least three bands of ß-globin amplified from DNA (n=115/135), and a RIN number ≥7 was considered very good for RNA (n=80/135). Sample preparation and storage time had little effect on RNA or DNA quality. Protein expression was assessed on tissue microarray by immunohistochemistry with antibodies against p53, WT1, E-cadherin, CK-7, and Ki67 from formalin fixed-paraffin embedded (FFPE) tissues. As seen with a nonhierarchical clustering statistical method, there was no significant difference in immunostaining of paraffin tissues among specimens from different biobanks. Interestingly, patients with worse outcome were highly positive for p53 and weak for WT1. In conclusion, while there was no common SOP for retrospectively collected material across Canadian biobanks, these results indicate that specimens collected at these multiple sites are of comparable quality, and can serve as an adequate resource to create a national cohort for the validation of molecular biomarkers in ovarian cancer.
1 Centre Hospitalier de l'Université de Montréal , Montreal, Quebec.
Cécile
C
Le Page
Martin
M
Köbel
Manon
M
de Ladurantaye
Kurosh
K
Rahimi
Jason
J
Madore
Sindy
S
Babinszky
Dimcho R
DR
Bachvarov
Magdalena
M
Bachvarova
Marie-Claude
MC
Beauchamp
Carol E
CE
Cass
Dianne
D
Chadwick
Crane
C
Colleen
Sambasivarao
S
Damaraju
Jennifer
J
Dufour
Walter H
WH
Gotlieb
Steve E
SE
Kalloger
Lise
L
Portelance
Jessica N
JN
McAlpine
Isabelle
I
Matte
Alain
A
Piché
Patricia
P
Shaw
Michael H A
MH
Roehrl
Barbara C
BC
Vanderhyden
Peter H
PH
Watson
David G
DG
Huntsman
Diane M
DM
Provencher
Anne-Marie
AM
Mes-Masson
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018070:Biological Specimen Banks / Q000592:standards*; D002170:Canada; D004273:DNA, Neoplasm / Q000378:metabolism; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000378:metabolism; D016612:Paraffin Embedding; D012334:RNA, Neoplasm / Q000378:metabolism; D013048:Specimen Handling / Q000592:standards*; D013194:Staining and Labeling; D046888:Tissue Array Analysis
D004273:DNA, Neoplasm; D012334:RNA, Neoplasm