Tong J, Gianni A M, Siena S, Srour E F, Bregni M, Hoffman R
Hematology/Oncology Section, Indiana University School of Medicine, Indianapolis, USA.
Blood Cells. 1994;20(2-3):351-62; discussion 362-3.
In this report we have used an in vitro assay for long-term culture-initiating cells (LTC-IC) to detect primitive hematopoietic progenitor cells (HPC) in the peripheral blood (PB) of cancer patients who received high-dose cyclophosphamide (HD-CTX) followed by a combination of recombinant hematopoietic growth factors (C-HGF) including either interleukin-3 (IL-3) + granulocyte colony-stimulating factor (G-CSF), IL-3 + granulocyte-macrophage colony-stimulating factor (GM-CSF) or IL-3/GM-CSF fusion protein (PIXY-321). In addition, we have developed a quantitative assay for cells capable of generating additional colony-forming cells (pre-CFC) as a means of determining primitive HPC present in mobilized PB cells. CD34+ human leukocyte A (HLA)-DR- cells isolated from the mobilized PB were capable of initiating long-term hematopoiesis in vitro that persisted for 10 weeks, while CD34+ HLA-DR- cells obtained from the nonmobilized PB or BM were capable of sustaining long-term hematopoiesis in vitro for only 4 weeks and 8 weeks, respectively. As determined by a limiting dilution analysis of mobilized PB CD34+ HLA-DR- cells, the frequency of pre-CFC was 4.3% (range, 1.0-8.3%). Pre-CFC comprised 0.01% (range, 0.001-0.02%) of mobilized PB mononuclear cells, and 151 pre-CFC were calculated to be present in one milliliter of mobilized PB (range, 20-310/ml). These results suggest that PB mononuclear cells collected by leukapheresis following mobilization with HD-CTX + C-HGFs contain not only differentiated HPCs but also more primitive HPC.
在本报告中，我们使用了一种针对长期培养起始细胞（LTC-IC）的体外检测方法，来检测接受大剂量环磷酰胺（HD-CTX）治疗后，再接受包括白细胞介素-3（IL-3）+粒细胞集落刺激因子（G-CSF）、IL-3+粒细胞-巨噬细胞集落刺激因子（GM-CSF）或IL-3/GM-CSF融合蛋白（PIXY-321）的重组造血生长因子联合治疗（C-HGF）的癌症患者外周血（PB）中的原始造血祖细胞（HPC）。此外，我们还开发了一种针对能够产生更多集落形成细胞的细胞（pre-CFC）的定量检测方法，以此作为确定动员的PB细胞中存在的原始HPC的手段。从动员的PB中分离出的CD34+人类白细胞A（HLA）-DR-细胞能够在体外启动持续10周的长期造血，而从未动员的PB或骨髓中获得的CD34+HLA-DR-细胞在体外仅能分别维持4周和8周的长期造血。通过对动员的PB CD34+HLA-DR-细胞进行有限稀释分析确定，pre-CFC的频率为4.3%（范围为1.0 - 8.3%）。pre-CFC占动员的PB单核细胞的0.01%（范围为0.001 - 0.02%），经计算每毫升动员的PB中存在151个pre-CFC（范围为20 - 310/ml）。这些结果表明，在HD-CTX + C-HGFs动员后通过白细胞分离术收集的PB单核细胞不仅含有分化的HPC，还含有更原始的HPC。
