Effects of cell cycle activation on the short-term engraftment properties of ex vivo expanded murine hematopoietic cells.
Effects of cell cycle activation on the short-term engraftment properties of ex vivo expanded murine hematopoietic cells.
复制标题
DOI：
10.1182/blood.v95.9.2829.009k37_2829_2837
复制
发表时间：
2000-05
期刊：
影响因子：
20.3
通讯作者：
S. Szilvassy;T. Meyerrose;B. Grimes
中科院分区：
文献类型：
--
作者：
S. Szilvassy;T. Meyerrose;B. Grimes
体外长期造血干细胞功能的丧失与细胞周期进展有关。为了确定细胞因子诱导的增殖是否也限制了体外扩增造血细胞的短期移植率和潜在的临床应用，我们将小鼠Sca-1(+)c-kit(+)Lin（-）细胞在白细胞介素-6 （IL-6）、IL-11、粒细胞集落刺激因子（G-CSF）、干细胞因子、flk-2配体和血小板生成素中培养7天。扩增2000倍的细胞用Hoechst 33342染色，通过流动分选分离成G(0)/G(1)（72% +/- 3%）或S/G(2)/M（27% +/- 3%）部分，注射到致死照射小鼠体内。虽然长期（超过6个月）淋巴和髓系移植在移植时的G(0)/G(1)扩增细胞的原发和继发受体中更大，但循环血细胞的短期（少于6周）恢复动力学没有明显差异。当造血细胞在含有四肽干细胞抑制剂n -乙酰基- ser - asp - lys - pro （AcSDKP）的培养物中扩增以减少移植前的祖细胞循环时，同样没有观察到受抑制或未受抑制的细胞在短期重建中的差异。有趣的是，AcSDKP在体内移植时显著加速了扩增的造血细胞的植入。大约1周后，白细胞恢复到正常水平的20%，acsdkp治疗的小鼠与未治疗的小鼠相比，血小板减少症基本消失。因此，虽然AcSDKP可以加速体外扩增的造血祖细胞的植入，这表明了一种相对简单的方法来提高其临床效用，但其作用似乎与细胞周期阻滞无关。(血。2000;95:2829 - 2837)
Loss of long-term hematopoietic stem cell function in vitro is associated with cell cycle progression. To determine whether cytokine-induced proliferation also limits the rate of short-term engraftment and potential clinical utility of ex vivo expanded hematopoietic cells, murine Sca-1(+)c-kit(+)Lin(-) cells were cultured in interleukin-6 (IL-6), IL-11, granulocyte colony-stimulating factor (G-CSF), stem cell factor, flk-2 ligand, and thrombopoietin for 7 days. Cells amplified 2000-fold were then stained with Hoechst 33342, separated into G(0)/G(1) (72% +/- 3%) or S/G(2)/M (27% +/- 3%) fractions by flow sorting, and injected into lethally irradiated mice. Although long-term (more than 6 months) engraftment of lymphoid and myeloid lineages was greater in primary and secondary recipients of expanded cells residing in G(0)/G(1) at the time of transplantation, there were no noted differences in the short-term (less than 6 weeks) recovery kinetics of circulating blood cells. When hematopoietic cells were expanded in cultures containing the tetrapeptide stem cell inhibitor N-Acetyl-Ser-Asp-Lys-Pro (AcSDKP) to reduce progenitor cycling prior to transplantation, again there were no differences observed in short-term reconstitution by inhibited or uninhibited cells. Interestingly, AcSDKP significantly accelerated engraftment by expanded hematopoietic cells when administered in vivo at the time of transplantation. Leukocytes recovered to 20% of normal levels approximately 1 week faster, and thrombocytopenia was largely abrogated in AcSDKP-treated versus untreated mice. Therefore, while AcSDKP can accelerate the engraftment of ex vivo expanded hematopoietic progenitors, which suggests a relatively simple approach to improve their clinical utility, its effects appear unrelated to cell cycle arrest. (Blood. 2000;95:2829-2837)