Department of Orthopaedic Surgery, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan.
J Neurosurg. 2012 Oct;117(4):787-94. doi: 10.3171/2012.7.JNS111503. Epub 2012 Aug 10.
CD133(+) cells have the potential to enhance histological and functional recovery from peripheral nerve injury. However, the number of CD133(+) cells safely obtained from human peripheral blood is extremely limited. To address this issue, the authors expanded CD133(+) cells derived from human peripheral blood using the serum-free expansion culture method and transplanted these ex vivo expanded cells into a model of sciatic nerve defect in rats. The purpose of this study was to determine the potential of ex vivo expanded CD133(+) cells to induce or enhance the repair of injured peripheral nerves.
Phosphate-buffered saline (PBS group [Group 1]), 10(5) fresh CD133(+) cells (fresh group [Group 2]), 10(5) ex vivo expanded CD133(+) cells (expansion group [Group 3]), or 10(4) fresh CD133(+) cells (low-dose group [Group 4]) embedded in atelocollagen gel were transplanted into a silicone tube that was then used to bridge a 15-mm defect in the sciatic nerve of athymic rats (10 animals per group). At 8 weeks postsurgery, histological and functional evaluations of the regenerated tissues were performed.
After 1 week of expansion culture, the number of cells increased 9.6 ± 3.3-fold. Based on the fluorescence-activated cell sorting analysis, it was demonstrated that the initial freshly isolated CD133(+) cell population contained 93.22% ± 0.30% CD133(+) cells and further confirmed that the expanded cells had a purity of 59.02% ± 1.58% CD133(+) cells. However, the histologically and functionally regenerated nerves bridging the defects were recognized in all rats in Groups 2 and 3 and in 6 of 10 rats in Group 4. The nerves did not regenerate to bridge the defect in any of the rats in Group 1.
The authors' results show that ex vivo expanded CD133(+) cells derived from human peripheral blood have a therapeutic potential similar to fresh CD133(+) cells for peripheral nerve injuries. The ex vivo procedure that can be used to expand CD133(+) cells without reducing their function represents a novel method for developing cell therapy for nerve defects in a clinical setting.
CD133(+)细胞具有增强周围神经损伤后组织学和功能恢复的潜力。然而，从人外周血中安全获得的 CD133(+)细胞数量极其有限。为了解决这个问题，作者使用无血清扩增培养方法扩增了来源于人外周血的 CD133(+)细胞，并将这些体外扩增的细胞移植到大鼠坐骨神经缺损模型中。本研究的目的是确定体外扩增的 CD133(+)细胞诱导或增强损伤周围神经修复的潜力。
磷酸盐缓冲盐水（PBS 组[第 1 组]）、10(5)个新鲜 CD133(+)细胞（新鲜组[第 2 组]）、10(5)个体外扩增的 CD133(+)细胞（扩增组[第 3 组]）或 10(4)个新鲜 CD133(+)细胞（低剂量组[第 4 组]）嵌入纤维连接蛋白凝胶中，然后移植到硅胶管中，硅胶管用于桥接去胸腺大鼠坐骨神经 15mm 的缺损（每组 10 只动物）。手术后 8 周，对再生组织进行组织学和功能评估。
经过 1 周的扩增培养，细胞数量增加了 9.6±3.3 倍。基于荧光激活细胞分选分析，证明初始分离的新鲜 CD133(+)细胞群体中含有 93.22%±0.30%的 CD133(+)细胞，并进一步证实扩增细胞的纯度为 59.02%±1.58%的 CD133(+)细胞。然而，在第 2 组和第 3 组的所有大鼠以及第 4 组的 10 只大鼠中的 6 只中，均识别到了桥接缺损的组织学和功能再生神经。在第 1 组的任何大鼠中，神经均未再生以桥接缺损。
作者的结果表明，来源于人外周血的体外扩增 CD133(+)细胞具有与新鲜 CD133(+)细胞相似的治疗周围神经损伤的潜力。这种可以在不降低其功能的情况下扩增 CD133(+)细胞的体外程序代表了一种在临床环境中开发神经缺陷细胞治疗的新方法。
