Baines P, Truran L, Bailey-Wood R, Hoy T, Lake H, Poynton C H, Burnett A
Department of Haematology, University Hospital of Wales, Cardiff.
Br J Haematol. 1994 Nov;88(3):472-80. doi: 10.1111/j.1365-2141.1994.tb05062.x.
We have measured the in vitro growth requirements of progenitor cells released into the blood of cancer patients following administration of chemotherapy and cytokines. In order to distinguish the direct effects of cytokines on progenitors from those activating accessory cells, we have compared clonogenic growth before and after CD34-positive selection of progenitors, in serum-free conditions. CD34 selection had little effect on the cytokine requirements of erythroid colony-forming cells and single cytokines, particularly interleukin-3, could support considerable colony growth in both mononuclear and CD34+ cell suspensions. Optimal erythroid colony growth, however, usually required the addition of a combination of stem cell factor and interleukin-3, in addition to erythropoietin, which was always required. Maximal numbers of granulocyte-monocyte progenitors in mononuclear cell cultures, could be achieved with a mixture of stem cell factor, interleukin-3 and granulocyte-monocyte colony stimulating factor. However, after CD34 selection, full myeloid colony growth was only achieved when granulocyte colony stimulating factor was added to the above mixture. This presumably reflects loss of accessory cells, during CD34 selection, which produce this cytokine. When transplanted after 8 d of culture, 16/22 myeloid colonies from erythropoietin-free cultures of peripheral blood stem cell harvests, could generate secondary erythroid colonies implying that these progenitors are multipotential. However, surface marker analysis of individual erythroid colonies revealed only the occasional presence of granulocytes and monocytes. These data demonstrate that cytokine mixtures are required for optimal colony growth, particularly after CD34 selection, and that most mobilized, blood clonogenic cells are multipotential.
我们已经测量了癌症患者在接受化疗和细胞因子治疗后释放到血液中的祖细胞的体外生长需求。为了区分细胞因子对祖细胞的直接作用与激活辅助细胞的作用，我们在无血清条件下比较了祖细胞CD34阳性选择前后的集落形成生长情况。CD34选择对红系集落形成细胞的细胞因子需求影响很小，单一细胞因子，特别是白细胞介素-3，可支持单核细胞和CD34+细胞悬液中大量的集落生长。然而，最佳的红系集落生长通常除了始终需要的促红细胞生成素外，还需要添加干细胞因子和白细胞介素-3的组合。单核细胞培养物中粒细胞-单核细胞祖细胞的最大数量，可通过干细胞因子、白细胞介素-3和粒细胞-单核细胞集落刺激因子的混合物实现。然而，在CD34选择后，只有在上述混合物中添加粒细胞集落刺激因子时，才能实现完全的髓系集落生长。这可能反映了在CD34选择过程中产生这种细胞因子的辅助细胞的丢失。在培养8天后移植时，外周血干细胞收获物的无促红细胞生成素培养物中的16/22个髓系集落可产生次级红系集落，这意味着这些祖细胞具有多能性。然而，对单个红系集落的表面标志物分析仅偶尔发现粒细胞和单核细胞的存在。这些数据表明，最佳的集落生长需要细胞因子混合物，特别是在CD34选择之后，并且大多数动员的血液集落形成细胞具有多能性。
