Stability of RNA isolated from human trabecular bone at post-mortem and surgery.
To determine the reliability of gene expression studies in human post-mortem bone, it is important to evaluate the stability of RNA isolated from such tissues as a function of the post-mortem interval. The stability of total RNA and bone-specific mRNA species was examined in bone samples obtained from routine autopsies and at surgery. The optimal temperature for any storage and transport of the bone before RNA isolation was shown to be 4 degrees C, and RT-PCR analysis is the preferred technique for the analysis of gene expression in post-mortem bone as it tolerates partial RNA degradation. For gene expression studies in bone, post-mortem cases, with a post-mortem interval of less than 48 h, should be selected, and the time that bone is stored after retrieval at autopsy or surgery should be kept to a minimum. Overall, our findings indicate that with appropriate storage and handling, RNA can be reliably isolated from human bone obtained at post-mortem and surgery to study ex vivo the pattern of gene expression in healthy individuals and in patients with musculoskeletal diseases such as osteoporosis and osteoarthritis.
Kuliwaba
Julia S
JS
Bone and Joint Research Laboratory, Division of Tissue Pathology, Institute of Medical and Veterinary Science and Hanson Institute, Frome Road, Adelaide 5000, South Australia, Australia. julia.kuliwaba@imvs.sa.gov.au
Fazzalari
Nicola L
NL
Findlay
David M
DM
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001344:Autopsy; D001842:Bone and Bones / Q000378:metabolism* / Q000601:surgery; D005260:Female; D015870:Gene Expression; D006801:Humans; D008297:Male; D008875:Middle Aged; D010003:Osteoarthritis / Q000235:genetics / Q000378:metabolism* / Q000601:surgery; D011180:Postmortem Changes; D012313:RNA / Q000302:isolation & purification*; D020871:RNA Stability; D012333:RNA, Messenger / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013048:Specimen Handling
D012333:RNA, Messenger; D012313:RNA