Holyoake T L, Freshney M G, McNair L, Parker A N, McKay P J, Steward W P, Fitzsimons E, Graham G J, Pragnell I B
Cancer Research Campaign Beatson Laboratories, Beaston Institute for Cancer Research, Glasgow, Scotland.
Blood. 1996 Jun 1;87(11):4589-95.
The characterization of many cytokines involved in the control of hematopoiesis has led to intense investigation into their potential use in ex vivo culture to expand progenitor numbers. We have established the optimum ex vivo culture conditions that allow substantial amplification of transient engrafting murine stem cells and which, simultaneously, augment the ability to sustain serial bone marrow transplantation (BMT). Short-term incubation of unfractionated BM cells in liquid culture with stem cell factor (SCF) and interleukin-11 (IL-11) produced a 50-fold amplification of clonogenic multipotential progenitors (CFU-A). Following such ex vivo expansion, substantially fewer cells were required to rescue lethally irradiated mice. When transplanted in cell doses above threshold for engraftment, BM cells expanded ex vivo resulted in significantly more rapid hematopoietic recovery. In a serial transplantation model, unmanipulated BM was only able to consistently sustain secondary BMT recipients, but BM expanded ex vivo has sustained quaternary BMT recipients that remain alive and well more than 140 days after 4th degree BMT. These results show augmentation of both short-term recovery posttransplant and the ability to serially transplant marrow by preincubation in culture with SCF and IL-11.
许多参与造血调控的细胞因子的特性，引发了对其在体外培养中用于扩增祖细胞数量的潜在用途的深入研究。我们已经建立了最佳的体外培养条件，该条件能够大量扩增短暂植入的小鼠干细胞，同时增强维持连续骨髓移植（BMT）的能力。将未分级的骨髓细胞与干细胞因子（SCF）和白细胞介素-11（IL-11）在液体培养基中短期孵育，可使克隆性多能祖细胞（CFU-A）扩增50倍。经过这种体外扩增后，拯救致死性照射小鼠所需的细胞数量大幅减少。当以高于植入阈值的细胞剂量进行移植时，体外扩增的骨髓细胞可导致造血恢复明显加快。在连续移植模型中，未处理的骨髓只能持续支持二次BMT受体，但体外扩增的骨髓已经支持了四级BMT受体，这些受体在第四次BMT后140多天仍存活且状况良好。这些结果表明，通过与SCF和IL-11在培养基中预孵育，可增强移植后的短期恢复能力以及连续移植骨髓的能力。
