Simpson David, Kakarala Gopikrishna, Hampson Karen, Steele Niall, Ashton Brian
Institute of Science and Technology in Medicine, University of Keele, the Robert Jones and Agnes Hunt Orthopaedic Hospital, Oswestry, Shropshire, UK.
Acta Orthop. 2007 Feb;78(1):26-30. doi: 10.1080/17453670610013385.
Fresh frozen bone allograft is available for human recipients after at least 6 months of quarantine at -80 degrees C. It is assumed that cryopreservation without cryoprotectant removes all viable donor cells.
We studied the in vitro cell growth from samples of fresh frozen human femoral head allografts after they had been released for patient use, and compared it with cell growth from a control group of fresh cancellous bone specimens from excised femoral heads (8 samples in each group).
Cell outgrowths were seen in all of the fresh cancellous bone specimens (100% of replicates, 48 replicates per specimen) but only in a small minority of replicates from 4 of the allograft samples (mean 3.1%). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) investigations revealed that cell outgrowths from both groups contained mRNA for transcription factors Runx2 and Osterix, and also for matrix proteins collagen type I, osteocalcin and bone sialoprotein. This is consistent with the cells being osteoblast-related.
This study confirms that fresh frozen human bone allograft cells have the potential to grow in vitro, but the significance of this in recipients is currently unknown.
新鲜冷冻骨同种异体移植物在-80℃下至少经过6个月的检疫后可供人类受者使用。假定无冷冻保护剂的冷冻保存可去除所有有活力的供体细胞。
我们研究了新鲜冷冻的人股骨头同种异体移植物样本在供患者使用后其体外细胞生长情况，并将其与来自切除股骨头的新鲜松质骨标本对照组的细胞生长情况进行比较（每组8个样本）。
所有新鲜松质骨标本均可见细胞生长（100%的重复样本，每个标本48个重复样本），但同种异体移植物样本中只有4个样本的一小部分重复样本可见细胞生长（平均3.1%）。逆转录定量聚合酶链反应（RT-qPCR）研究显示，两组的细胞生长物均含有转录因子Runx2和Osterix以及基质蛋白I型胶原、骨钙素和骨唾液蛋白的mRNA。这与细胞为成骨细胞相关一致。
本研究证实新鲜冷冻的人骨同种异体移植物细胞具有体外生长的潜力，但这对受者的意义目前尚不清楚。
