Ottmann O G, Stella C C, Eder M, Reutzel P, Ströcker S, Hoelzer D, Ganser A
Department of Hematology, University of Frankfurt, FRG.
Exp Hematol. 1991 Sep;19(8):773-8.
Our experiments have addressed the regulation of early hematopoietic progenitor cell expansion by interleukin 3 (IL-3) and interleukin 1 beta (IL-1 beta) and its modulation by bone marrow fibroblasts in vitro. In a two-stage assay utilizing serum-deprived (SD) presuspension cultures of CD34-enriched bone marrow (BM) cells followed by clonal cultures, absolute numbers of granulocyte-macrophage progenitor cells (day-14 granulocyte-macrophage colony-forming units [CFU-GM]) increased progressively to 164% and 204% of input levels after 12 days of culture in the presence of IL-3 alone or in combination with IL-1 beta, respectively. Multilineage (granulocyte erythrocyte macrophage megakaryocyte colony-forming units, CFU-GEMM) and erythroid (erythroid burst-forming units, BFU-E) progenitor cell numbers increased above or were maintained at input levels after 4 and 7 days of liquid culture in the presence of IL-3 and IL-3 plus IL-1 beta, respectively, but in contrast to granulocyte-macrophage colony-forming units (CFU-GM) they were essentially undetectable after 12 days of culture. Progenitors more primitive than colony-forming cells (pre-CFU) were assessed in SD-presuspension cultures of CD34-enriched BM cells purged with mafosfamide to eliminate base-line CFU-GM, CFU-GEMM, and BFU-E. Under these conditions and in the absence of stromal elements, CFU-GM but neither CFU-GEMM nor BFU-E developed in response to cytokines alone. In the additional presence of passaged bone marrow fibroblasts, however, IL-3 plus IL-1 beta and to a lesser degree IL-3 alone induced a pronounced amplification of BFU-E and CFU-GEMM, indicating that their development from a more primitive progenitor compartment requires growth activities in addition to IL-3 and IL-1 beta that are provided by marrow-derived stromal cells such as fibroblasts.
我们的实验研究了白细胞介素3（IL-3）和白细胞介素1β（IL-1β）对早期造血祖细胞扩增的调节作用，以及骨髓成纤维细胞在体外对其的调控。在一个两阶段试验中，先对富含CD34的骨髓（BM）细胞进行血清饥饿（SD）预悬浮培养，然后进行克隆培养。在单独使用IL-3或与IL-1β联合培养12天后，粒细胞-巨噬细胞祖细胞的绝对数量（第14天的粒细胞-巨噬细胞集落形成单位[CFU-GM]）分别逐渐增加至输入水平的164%和204%。多系（粒细胞-红细胞-巨噬细胞-巨核细胞集落形成单位，CFU-GEMM）和红系（红系爆式集落形成单位，BFU-E）祖细胞数量在分别用IL-3和IL-3加IL-1β进行液体培养4天和7天后增加到输入水平以上或维持在输入水平，但与粒细胞-巨噬细胞集落形成单位（CFU-GM）不同，培养12天后它们基本检测不到。在对富含CD34的BM细胞进行SD预悬浮培养并用马磷酰胺清除基线CFU-GM、CFU-GEMM和BFU-E后，评估比集落形成细胞更原始的祖细胞（前CFU）。在这些条件下且无基质成分时，单独的细胞因子可使CFU-GM发育，但CFU-GEMM和BFU-E均不发育。然而，在加入传代骨髓成纤维细胞后，IL-3加IL-1β以及单独的IL-3在较小程度上可诱导BFU-E和CFU-GEMM显著扩增，这表明它们从更原始的祖细胞区室发育除了需要IL-3和IL-1β外，还需要骨髓来源的基质细胞如成纤维细胞提供的生长活性。
