Transient production of bone morphogenetic protein 2 by allogeneic transplanted transduced cells induces bone formation
Transient production of bone morphogenetic protein 2 by allogeneic transplanted transduced cells induces bone formation
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DOI：
10.1089/10430340050016274
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发表时间：
2000-01-01
影响因子：
4.2
通讯作者：
Lyons, KM
中科院分区：
文献类型：
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作者：
Engstrand, T;Daluiski, A;Lyons, KM
本研究的目的是评估使用基因修饰的同种异体细胞移植作为诱导骨形成的方法。在这项研究中，我们用含有人骨形态发生蛋白2（BMP 2）基因的逆转录病毒载体感染小鼠骨祖细胞系，转导的细胞比任何测试剂量的重组人BMP 2蛋白（rhBMP 2）处理的细胞表现出更高的碱性磷酸酶活性。将转导的细胞悬浮在胶原蛋白溶液中，并注射到免疫活性远系小鼠的四头肌中。放射学和组织学检查显示，85%的移植动物（n = 13）中有丰富的异位骨形成，嘌呤霉素抗性基因的PCR和Southern印迹分析显示，移植细胞可检测到长达1周，但在以后的时间点不可检测。所有动物均未发生肿瘤。我们的研究结果表明，同种异体BMP 2表达转导细胞可能具有治疗潜力，促进新骨形成。该模型还提供了一种简单、廉价和灵敏的测定方法，用于在体内评价分泌蛋白的骨诱导潜力，而不需要蛋白纯化或使用免疫缺陷动物。
The aim of this study was to evaluate the use of transplantation of genetically modified allogeneic cells as a method to induce bone formation. In this study, we infected a murine osteoprogenitor cell line with a retroviral vector containing the human bone morphogenic protein 2 (BMP2) gene, Transduced cells exhibited more alkaline phosphatase activity than cells treated with any of the tested doses of recombinant human BMP2 protein (rhBMP2). The transduced cells were suspended in a collagen solution and injected into the quadriceps muscle in immunocompetent outbred mice. Radiographic and histological examinations demonstrate abundant ectopic bone formation in 85% of the transplanted animals (n = 13), PCR and Southern blot analysis for the puromycin resistance gene revealed that the transplanted cells were detectable for up to 1 week, but not at later time points. None of the animals developed tumors. Our results suggest that allogeneic BMP2 expressing transduced cells may have therapeutic potential for enhancing new bone formation. This model also provides a simple, inexpensive, and sensitive assay for evaluating in vivo the osteoinductive potentials of secreted proteins without the requirement of protein purification or the use of immunodeficient animals.