EFFECT OF STEM-CELL FACTOR (C-KIT LIGAND), GRANULOCYTE-MACROPHAGE COLONY-STIMULATING FACTOR AND INTERLEUKIN-3 ON HEMATOPOIETIC PROGENITORS IN HUMAN LONG-TERM BONE-MARROW CULTURES
EFFECT OF STEM-CELL FACTOR (C-KIT LIGAND), GRANULOCYTE-MACROPHAGE COLONY-STIMULATING FACTOR AND INTERLEUKIN-3 ON HEMATOPOIETIC PROGENITORS IN HUMAN LONG-TERM BONE-MARROW CULTURES
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DOI：
10.1002/stem.5530110511
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发表时间：
1993-09-01
期刊：
影响因子：
5.2
通讯作者：
GULATI, SC
中科院分区：
文献类型：
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作者：
LEMOLI, RM;GULATI, SC
本文试图对造血干细胞扩增方法进行改进。我们评估了重组人干细胞因子（SCF或c-kit配体）单独使用以及与重组人粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白细胞介素3 （IL-3）联合使用对人长期骨髓培养（LTBMC）细胞增殖和分化的影响。每周添加5 ng/ml含25%血清培养基的SCF可在第1周增加总非贴壁细胞、粒细胞-巨噬细胞集落形成单位（CFU-GM）和红细胞爆发形成单位（BFU-E）的恢复，但骨髓（BM）祖细胞的数量在第3周（BFU-E）和第4周（CFU-GM）低于未处理对照培养的水平。在第8周，当终止培养时，与对照组相比，添加SCF的烧瓶中CFU-GM恢复明显减少（p < 0.002）。此外，SCF治疗可导致BFU-E早期消失。当LTBMC中添加SCF + GM-CSF （100 U/ml）和IL-3 （5 ng/ml）时，在第1周观察到csf的协同活性。当SCF与GM-CSF联合使用时，BM集落形成细胞（CFC）的数量迅速下降到低于无生长因子对照的水平，导致培养物早期衰竭。通过比较，GM-CSF和IL-3单独诱导CFU-GM和BFU-E的非贴壁细胞菌落数量或非贴壁细胞菌落数量高于对照组（无生长因子），具有统计学意义。对添加SCF培养物的贴壁层细胞的分析显示，细胞数量增加，没有脂肪生成，骨髓祖细胞早期消失，而通过阿拉伯糖胞苷（Ara-C）自杀试验评估的S期CFU-GM的百分比为9.2 +/- 5% SD，而对照组（无生长因子）样品的百分比为27.7 +/- 10% SD （p < 0.01）。SCF增加了成纤维细胞集落形成单位（CFU-F）的数量，并与IL-3联合使用时显示出协同作用（增加9.6倍）。这些发现表明SCF、GM-CSF和IL-3在造血系统内不同的细胞群中发挥其活性。需要进一步的研究来优化SCF在支持造血中的应用。
In this paper we attempt to improve upon the methods of hematopoietic stem cell expansion. We evaluate the effects of recombinant human stem cell factor (SCF or c-kit ligand) alone and also in combination with recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin 3 (IL-3), on cell proliferation and differentiation in human long term bone marrow cultures (LTBMC). Weekly addition of 5 ng/ml of SCF with 25% serum containing media resulted in increased recovery of total nonadherent cells, granulocyte-macrophage colony forming units (CFU-GM), and burst-forming units erythroid (BFU-E) at week 1, but the number of bone marrow (BM) progenitor cells fell below the level of untreated control cultures at weeks 3 (BFU-E) and 4 (CFU-GM). At week 8, when the cultures were terminated, the CFU-GM recovery was markedly reduced in flasks supplemented with SCF compared with the controls (p < 0.002). Moreover, SCF treatment induced the early disappearance of BFU-E. When LTBMC were supplemented with the combination of SCF plus GM-CSF (100 U/ml) and IL-3 (5 ng/ml), synergistic activity of the CSFs was observed at week 1. The number of BM colony forming cells (CFC) rapidly declined below the level of growth factor-free controls, leading to the early exhaustion of the culture when SCF was combined with GM-CSF. By comparison, GM-CSF and IL-3 alone induced a statistically significant increase above the controls (no growth factor) in the number or nonadherent cell colonies of CFU-GM and BFU-E. Analysis of adherent layer cells from cultures supplemented with SCF showed increased cellularity, no adipogenesis, and early disappearance of myeloid progenitors while the percentage of CFU-GM in S phase, assessed by cytosine arabinoside (Ara-C) suicide assay, was 9.2 +/- 5% SD versus 27.7 +/- 10% SD in control (no growth factor) samples (p < 0.01). SCF increased the number of fibroblast colony forming units (CFU-F) and also showed a synergistic activity (9.6-fold increase) when combined with IL-3. These findings suggest that SCF, GM-CSF and IL-3 exert their activity on different cell populations within the hematopoietic system. Further investigations are needed to optimize the use of SCF in supporting hematopoiesis.