Kanbara Tamotsu, Kurobe Hirotsugu, Kitaichi Takashi, Sugano Mikio, Nakayama Taisuke, Kinoshita Hajime, Iwase Takashi, Akaike Masafumi, Abe Masahiro, Sata Masataka, Matsumoto Toshio, Kitagawa Tetsuya
Department of Cardiovascular Surgery, Institute for Health Biosciences, The University of Tokushima Graduate School, Tokushima, Tokushima, Japan.
Ann Vasc Dis. 2012;5(1):52-60. doi: 10.3400/avd.oa.11.00070. Epub 2012 Feb 15.
Efficient and secure collection of CD34+ cells are crucial for the angiogenic therapies. We have developed autologous peripheral blood-mononuclear cell (MNC) transplantation induced by erythropoietin (rhEPO) for critical ischemic limbs.
Seven patients, including five with arteriosclerosis obliterans, one with Buerger's disease and one with progressive systemic sclerosis, underwent ten cell therapies. The first administration of rhEPO was performed two weeks before apheresis, and the second administration and blood donation were performed one week before apheresis to activate bone marrow. MNCs including CD34+ cells, isolated from peripheral blood by apheresis, were immediately injected intramuscularly into ischemic limbs.
The number of peripheral blood-CD34 + cells had significantly increased from 1.32 ± 0.83/microL, before the rhEPO induction, to 1.86 ± 0.94/microL, before the apheresis. The number of transplanted MNCs ranged between 0.5 × 10(9) and 16.5 × 10(9), and that of CD34+ cells, between 0.1 × 10(6) and 12.7 × 10(6), accounting for 0.02%-0.1% of MNCs. There were no serious complications. Finger ulcers with Buerger's disease were significantly improved one month after the transplantations, but the same or other ulcer(s) appeared 2-6 months later. Three patients had an improvement in rest pain, and one patient extended maximum pain-free walking distance.
Erythropoietin-induced autologous peripheral blood-MNC transplantation is a useful and safe alternative for ischemic limbs.
高效且安全地采集CD34+细胞对于血管生成疗法至关重要。我们已研发出用于严重缺血肢体的促红细胞生成素（rhEPO）诱导的自体外周血单个核细胞（MNC）移植方法。
7例患者接受了10次细胞治疗，其中5例为闭塞性动脉硬化症患者，1例为血栓闭塞性脉管炎患者，1例为进行性系统性硬化症患者。在进行单采术两周前首次给予rhEPO，在单采术前一周进行第二次给药及献血以激活骨髓。通过单采术从外周血中分离出包括CD34+细胞在内的MNC，并立即将其肌肉注射到缺血肢体中。
外周血CD34 +细胞数量从rhEPO诱导前的1.32±0.83/微升显著增加至单采术前的1.86±0.94/微升。移植的MNC数量在0.5×10(9)至16.5×10(9)之间，CD34+细胞数量在0.1×10(6)至12.7×10(6)之间，占MNC的0.02%-0.1%。未出现严重并发症。血栓闭塞性脉管炎患者的手指溃疡在移植后1个月有显著改善，但在2-6个月后又出现相同或其他溃疡。3例患者静息痛有所改善，1例患者最大无痛行走距离延长。
促红细胞生成素诱导的自体外周血MNC移植是缺血肢体一种有用且安全的替代疗法。
