Williams S F, Lee W J, Bender J G, Zimmerman T, Swinney P, Blake M, Carreon J, Schilling M, Smith S, Williams D E, Oldham F, Van Epps D
Department of Medicine, University of Chicago Medical Center, IL, USA.
Blood. 1996 Mar 1;87(5):1687-91.
Cytopenia after high-dose chemotherapy and autologous stem cell reinfusion is a major cause of morbidity. Ex vivo cultured expansion and differentiation of CD34+ peripheral blood progenitor cells (PBPC) to neutrophil precursors may shorten the neutropenic period further. We explored the use of these ex vivo cultured PBPCs in nine patients with metastatic breast cancer. All underwent PBPC mobilization with cyclophosphamide, VP-16, and G-CSF. Subsequently, they underwent four to five apheresis procedures. One apheresis product from each patient was prepared using the Isolex 300 Magnetic Cell Separation System (Baxter Immunotherapy, Irvine, CA) to obtain CD34+ cells. These cells were then cultured in gas permeable bags containing serum-free X-VIVO 10 (BioWhittaker, Walkersville, MD) medium supplemented with 1% human serum albumin and 100 ng/mL PIXY321. At day 12 of culture the mean fold expansion was 26x with a range of 6 to 64x. One patient's cells did not expand because of a technical difficulty. The final cell product contained an average of 29.3% CD15+ neutrophil precursors with a range of 18.5% to 48.1%. The patients underwent high-dose chemotherapy with cyclophosphamide, carboplatin, and thiotepa. On day 0, the cryopreserved PBPCs were reinfused and on day +1 the 12-day cultured cells were washed, resuspended, and reinfused into eight of nine patients. One patient was not infused with cultured cells. The mean number of cultured cells reinfused was 44.6 x 10(6) cells/kg with a range of 0.8 to 156.6 x 10(6) cells/kg. No toxicity was observed after reinfusion. The eight patients have recovered absolute neutrophil counts > 500/microL on a median of 8 days (range 8 to 10 days); the median platelet transfusion independence occurred on day 10 (range 8 to 12 days) and platelet counts > 50,000/microL were achieved by day 12 (range 9 to 14) for the seven patients whose platelet counts could be determined. Expanded CD34+ selected PBPC can be obtained and safely reinfused into patients.
大剂量化疗及自体干细胞回输后的血细胞减少是发病的主要原因。将CD34 +外周血祖细胞（PBPC）体外培养扩增并分化为中性粒细胞前体可能会进一步缩短中性粒细胞减少期。我们在9例转移性乳腺癌患者中探索了使用这些体外培养的PBPC。所有患者均接受环磷酰胺、VP - 16和G - CSF进行PBPC动员。随后，他们接受了4至5次单采程序。使用Isolex 300磁性细胞分离系统（百特免疫治疗公司，加利福尼亚州欧文市）制备每位患者的一份单采产物以获得CD34 +细胞。然后将这些细胞在含有无血清X - VIVO 10（BioWhittaker公司，马里兰州沃克维尔）培养基）培养基中培养，该培养基补充有1%人血清白蛋白和100 ng/mL PIXY321。培养第12天时，平均扩增倍数为26倍，范围为6至64倍。1例患者的细胞因技术问题未扩增。最终细胞产物平均含有29.3%的CD15 +中性粒细胞前体，范围为18.5%至48.1%。患者接受了环磷酰胺、卡铂和噻替哌的大剂量化疗。第0天，回输冷冻保存的PBPC，第 +1天，将培养12天的细胞洗涤、重悬并回输给9例患者中的8例。1例患者未输注培养细胞。回输的培养细胞平均数量为44.6×10(6)个细胞/kg，范围为0.8至156.6×10(6)个细胞/kg。回输后未观察到毒性。8例患者的绝对中性粒细胞计数中位数在8天（范围8至10天）时恢复至> 500/μL；7例血小板计数可测定的患者血小板输注独立性中位数出现在第10天（范围8至12天），第12天（范围9至14天）时血小板计数> 50,000/μL。可以获得扩增的CD34 +选择的PBPC并安全地回输给患者。
