Effect of tissue shipping on plasma cell isolation, viability, and RNA integrity in the context of a centralized good laboratory practice-certified tissue banking facility.
The Multiple Myeloma Research Consortium has established a tissue bank for the deposition of bone marrow samples from patients with multiple myeloma to be mailed and processed under good laboratory practices. To date, over 1,000 samples have been collected. At this time, limited information is available on shipped bone marrow aspirates in regards to cell viability, yield, purity, and subsequent RNA yield and quality. To test these determinants, we did a pilot study on behalf of the Multiple Myeloma Research Consortium where samples were drawn at Mayo Clinic Rochester (MCR) pooled and split into two equal aliquots. One-half of each sample was processed following good laboratory practices compliant standard operating procedures, immediately after sample procurement, at MCR. The CD138+ cells were stored at -80 degrees C as a Trizol lysate. The other half of the aspirate was sent overnight to Mayo Clinic Scottsdale where they were processed using identical standard operating procedures. The RNA was extracted and analyzed in a single batch at MCR. At both locations, samples were assayed for the following quality determinants: Viability was assessed using a three-color flow cytometric method (CD45, CD38, and 7-AAD). Cell counts were done to determine plasma cell recovery and post-sort purity determined by means of a slide-based immunofluorescent assay. RNA recovery and integrity was assessed using the Agilent Bioanalyzer. Lastly, gene expression profiles were compared to determine the signature emanating from the shipment of samples. Despite minor differences, our results suggest that shipment of samples did not significantly affect these quality determinants in aggregate.
Ahmann
Gregory J
GJ
Comprehensive Cancer Center and Division of Hematology and Oncology, Mayo Clinic Arizona, Scottsdale, Arizona 85259, USA.
Chng
Wee Joo
WJ
Henderson
Kimberly J
KJ
Price-Troska
Tammy L
TL
DeGoey
Roberta W
RW
Timm
Michael M
MM
Dispenzieri
Angela
A
Greipp
Philip R
PR
Sable-Hunt
Alicia
A
Bergsagel
Leif
L
Fonseca
Rafael
R
D000704:Analysis of Variance; D001854:Bone Marrow Cells; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D020869:Gene Expression Profiling; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D010950:Plasma Cells; D012313:RNA / Q000032:analysis; D013048:Specimen Handling / Q000592:standards*; D014015:Tissue Banks
D012313:RNA