Homing of purified murine lymphohematopoietic stem cells: A cytokine-induced defect
Homing of purified murine lymphohematopoietic stem cells: A cytokine-induced defect
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DOI：
10.1089/152581602321080574
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发表时间：
2002-12-01
期刊：
影响因子：
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通讯作者：
Quesenberry, P
中科院分区：
文献类型：
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作者：
Cerny, J;Dooner, M;Quesenberry, P
本研究旨在用纯化的谱系阴性的SCA-1阳性(LIN(-)SCA(+))小鼠骨髓细胞建立直接归巢实验，并评价细胞因子对归巢的影响。用5-(和6)-羧基荧光素二乙酸酯琥珀酰亚胺酯(CFDA-SE)标记C57BL/6LIN(-)SCA(+)骨髓干细胞，尾静脉注射未处理的C57BL/6小鼠。在细胞输注后的不同时间收集骨髓，并在高速MoFlo细胞分选器上分析荧光阳性事件，使用大事件分析，分析了至少1600万个总事件。我们已经证明，LIN(-)SCA(+)细胞在输注后1h和3h的归巢在50,000到1,000,000个输注细胞之间是线性的。这为归巢试验奠定了基础，在归巢试验中，输入250,000个标记LIN(-)SCA(+)的骨髓细胞，然后在3小时后从骨髓中回收细胞，随后进行大事件荧光激活细胞分选(FACS)分析。我们发现7.45-9.32%的输注细胞归巢，培养48h的干细胞在白介素3(IL-3)、IL-6、IL-11和钢铁因子培养细胞中的归巢与未培养细胞相比存在缺陷。骨髓干细胞暴露于IL-3、IL-6、IL-11和钢铁因子会导致干细胞归巢缺陷，这可能是先前在这些条件下表现出的植入缺陷的基础。
This study was designed to establish a direct homing assay using purified lineage-negative Sca-1-positive (Lin(-) Sca(+)) murine bone marrow cells and to evaluate the effects of cytokines on, homing. C57BL/6 Lin(-) Sca(+) marrow stem cells were labeled with 5-(and 6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and then injected by tail vein into untreated C57BL/6 mice. Marrow was harvested at various times after cell infusion and analyzed on a high-speed MoFlo cell sorter for fluorescent positive events, using a large event analysis, with at least 16 million total events analyzed. We have shown that homing of Lin(-) Sca(+) cells plateaus by 1 h, and at 3 h post-infusion is linear between 50,000 and 1,000,000 infused cells. This forms a base for a homing assay in which 250,000 CFDA-SE labeled Lin(-) Sca(+) marrow cells are infused and then recovered from marrow 3 h later, followed by a large-event fluorescence-activated cell sorting (FACS) analysis. We found that 7.45-9.32% of infused cells homed and that homing of stem cells cultured for 48 h in interleukin-3 (IL-3), IL-6, IL-11, and steel factor cultured cells was defective when compared to noncultured cells. Exposure of marrow stem cells to IL-3, IL-6, IL-11, and steel factor induces a stem cell homing defect, which probably underlies the engraftment defect previously characterized under these conditions.