Heyligers Ide C, Klein-Nulend Jenneke
Department of Orthopaedic Surgery, Skeletal Tissue Engineering Group, Amsterdam (STEGA), Atrium MC, Heerlen, The Netherlands.
Cell Tissue Bank. 2005;6(1):25-31. doi: 10.1007/s10561-005-1089-4.
Impacted morselized donor bone is successfully used to treat bone loss in revision total hip arthroplasties. It is generally thought, but not proven, that the processing and storage at -80 degrees C of the donor bone kills all cells. Because of the risk of contamination and to increase our understanding about the process of new bone formation after revision total hip arthroplasty, the aim of this study was to investigate whether the donor bone does contain vital cells. Samples from 11 femoral heads were obtained according to the American and European standards of bone banking, and tested for their capacity to give rise to proliferating cells, using tissue culture methods. All bone samples were stored at - 80 degrees C for a minimum of 6 months. Bone sample cores were morselized and cultured for 6 weeks. Inverted phase contrast microscopy was used to evaluate cell growth. DNA marker analysis was used to confirm cellular identity. All bank bone samples gave rise to cell growth. The cell cultures showed osteoblastic characteristics in that they expressed high levels of alkaline phosphatase activity. DNA marker analysis showed identical alleles for cultured cells from frozen bone and freshly obtained buccal cells from the same donor, indicating that the cells growing from the banked bone were indeed originating from the donor tissue. It was therefore concluded that -80 degrees C freezing of bone tissue does not routinely kill cells within the tissue.
嵌压式碎骨移植供体骨成功用于治疗翻修全髋关节置换术中的骨缺损。人们普遍认为（但未经证实），供体骨在-80℃下进行处理和储存会杀死所有细胞。由于存在污染风险，也为了增进我们对翻修全髋关节置换术后新骨形成过程的了解，本研究的目的是调查供体骨是否确实含有活细胞。根据美国和欧洲骨库标准获取了11个股骨头的样本，并采用组织培养方法检测其产生增殖细胞的能力。所有骨样本均在-80℃下储存至少6个月。将骨样本芯磨碎并培养6周。使用倒置相差显微镜评估细胞生长情况。采用DNA标记分析来确认细胞身份。所有骨库样本均能产生细胞生长。细胞培养显示出成骨细胞特征，即它们表达高水平的碱性磷酸酶活性。DNA标记分析显示，来自冷冻骨培养的细胞与来自同一供体的新鲜获取的颊细胞具有相同的等位基因，这表明从骨库骨中生长的细胞确实源自供体组织。因此得出结论，骨组织在-80℃下冷冻通常不会杀死组织内的细胞。
