Uchida N, He D, Friera A M, Reitsma M, Sasaki D, Chen B, Tsukamoto A
Systemix Inc, Palo Alto, CA 94304, USA.
Blood. 1997 Jan 15;89(2):465-72.
Treatment with a combination of cytokines and chemotherapy can effectively stimulate the release of hematopoietic stem cells (HSC) into the peripheral blood (PB), which can then be harvested for transplantation. The cell cycle status of the harvested HSC from mobilized PB (MPB) is of interest because of the impact that cell cycling may have on optimizing the conditions for ex vivo expansion, retrovirus-mediated gene transfer, and the engraftment of transplanted tissues. Therefore, we characterized the cell cycling status of mobilized HSC from mice and humans. The murine HSC, which express the phenotype c-kit+ Thy-1.1lo Lin-/lo Sca-1+, were purified from PB, bone marrow (BM), and spleen after the mice were treated with the mobilizing regimen of granulocyte colony-stimulating factor (G-CSF) or a combination of cyclophosphamide (CTX) and G-CSF. Human HSC (CD34+ Thy-1+ Lin-) and progenitor cells (CD34+ Thy-1-Lin-) were isolated from the BM of untreated healthy volunteers and from MPB of healthy volunteers and patients treated with G-CSF or a combination of CTX and GM-CSF. Cell cycle status was determined by quantitating the amount of DNA in the purified cells after staining with the dye Hoechst 33342. Fluorescence-activated cell sorting analysis of the progenitor cells from the murine and human samples showed an unexpected finding, ie, virtually none of the cells from the MPB was cycling. The G0/G1 status of HSC from MPB was surprising, because a significant proportion of HSC from BM are actively proliferating and, after mobilization, the HSC in the spleen and BM were also actively cycling.
细胞因子与化疗联合治疗可有效刺激造血干细胞（HSC）释放到外周血（PB）中，随后可采集用于移植。由于细胞周期可能对优化体外扩增、逆转录病毒介导的基因转移以及移植组织的植入条件产生影响，因此来自动员外周血（MPB）的采集HSC的细胞周期状态备受关注。因此，我们对来自小鼠和人类的动员HSC的细胞周期状态进行了表征。在小鼠接受粒细胞集落刺激因子（G-CSF）或环磷酰胺（CTX）与G-CSF联合动员方案治疗后，从PB、骨髓（BM）和脾脏中纯化出表达c-kit+ Thy-1.1lo Lin-/lo Sca-1+表型的小鼠HSC。从未经治疗的健康志愿者的BM以及接受G-CSF或CTX与GM-CSF联合治疗的健康志愿者和患者的MPB中分离出人HSC（CD34+ Thy-1+ Lin-）和祖细胞（CD34+ Thy-1-Lin-）。通过用Hoechst 33342染料染色后定量纯化细胞中的DNA量来确定细胞周期状态。对小鼠和人类样本中的祖细胞进行荧光激活细胞分选分析发现了一个意外的结果，即MPB中的细胞几乎没有处于细胞周期中。MPB中HSC的G0/G1状态令人惊讶，因为BM中相当一部分HSC在活跃增殖，并且动员后，脾脏和BM中的HSC也在活跃地进行细胞周期循环。
