Li Yang, Wang Xiao-Ping, Guo Hai-Xia, Huang Ke, Wei Jing, Zhou Dun-Hua, Huang Wen-Ge, Wu Yan-Feng, Huang Shao-Liang
Pediatric Department, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, Guangdong Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Feb;18(1):140-5.
The study was aimed to explore the distribution and interaction mechanism of human bone marrow mesenchymal stem cells (MSC) and cord blood cytokine-induced killer (CIK)/natural killer (NK) cells infused via different ways at different times in NOD/SCID mice. 5 microl 1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine perchlorate (DiI) dye(red) was added in suspension of MSC per ml, and 1 microl carboxyfluorescein diacetate, succinimidyl ester(CFDA SE) dye(green) was added in suspension of CIK/NK cells per ml. The amounts of MSC and CIK/NK cells infused in each 6 NOD/SCID mouse were 1 x 10(6) (0.1 ml) and 1 x 10(7) (0.1 ml) respectively. All mice were divided into 4 groups, each group consisted of 6 mice. Group A: MSC (intravenous infusion, iv) + CIK/NK cells (iv) at the same time, group B: MSC (iv) + CIK/NK cells (iv) at 48 hours after infusion of MSC; group C: MSC (intramedullary infusion, im) + CIK/NK cells (iv) at the same time; group D: MSC (im) + CIK/NK cells (iv) at 48 hours after infusion of MSC. 3 NOD/SCID mice were sacrificed per batch at 24 hours and 48 hours after infused CIK/NK cells. Frozen sections of liver, spleen, lung and kidney were prepared, and then followed by counting the amounts of red and green fluorescence cells under fluorescence microscope, and calculating the ratio of MSC to CIK/NK cells for reflecting the interaction of MSC and CIK/NK cells in mice, and for showing the suppressive intensity of MSCs on CIK/NK cells. The results showed that the sums of average ratios of MSC to CIK/NK cells in lung, liver and spleen of group A and B were higher than that in group C and D at 24 hours and 48 hours respectively after infusing CIK/NK cells. The sum of average ratios of MSC to CIK/NK cells in group A was slightly higher than that in group B at 24 hours and 48 hours after infusing CIK/NK cells, but there was no significant difference between them. The sum of average ratios of MSC to CIK/NK cells in lung, liver and spleen in group C was slightly lower than that in group D at 24 hours after infusing CIK/NK cells, but reversed at 48 hours later and there was no significant difference between them. The sums of average ratios of MSC to CIK/NK cells in lung, liver and spleen in group A, B, C and D were all higher than those in kidney at 24 and 48 hours respectively after infusing CIK/NK cells. It is concluded, the MSC and CIK/NK cells may interact if they are infused via the same way and at the same time, the location where the suppression of MSC on CIK/NK cells occur in vivo may be reticulo-endothelial systems in lungs and livers.
本研究旨在探讨人骨髓间充质干细胞（MSC）和脐血细胞因子诱导的杀伤细胞（CIK）/自然杀伤细胞（NK）在不同时间以不同方式注入NOD/SCID小鼠后，其分布及相互作用机制。每毫升MSC悬液中加入5微升1,1'-二辛基-3,3,3',3'-四甲基吲哚羰花青高氯酸盐（DiI）染料（红色），每毫升CIK/NK细胞悬液中加入1微升羧基荧光素二乙酸琥珀酰亚胺酯（CFDA SE）染料（绿色）。每只6只NOD/SCID小鼠注入的MSC和CIK/NK细胞数量分别为1×10⁶（0.1毫升）和1×10⁷（0.1毫升）。所有小鼠分为4组，每组6只。A组：同时静脉注射（iv）MSC + CIK/NK细胞；B组：注入MSC 48小时后静脉注射CIK/NK细胞；C组：同时骨髓内注射（im）MSC + CIK/NK细胞；D组：注入MSC 48小时后静脉注射CIK/NK细胞。在注入CIK/NK细胞后24小时和48小时，每批处死3只NOD/SCID小鼠。制备肝、脾、肺和肾的冰冻切片，然后在荧光显微镜下计数红色和绿色荧光细胞数量，计算MSC与CIK/NK细胞的比例，以反映小鼠体内MSC与CIK/NK细胞的相互作用，并显示MSC对CIK/NK细胞的抑制强度。结果显示，在注入CIK/NK细胞后24小时和48小时，A组和B组肺、肝和脾中MSC与CIK/NK细胞平均比例之和分别高于C组和D组。在注入CIK/NK细胞后24小时和48小时，A组中MSC与CIK/NK细胞平均比例之和略高于B组，但两者之间无显著差异。在注入CIK/NK细胞后24小时，C组肺、肝和脾中MSC与CIK/NK细胞平均比例之和略低于D组，但48小时后相反，且两者之间无显著差异。在注入CIK/NK细胞后24小时和48小时，A、B、C、D组肺、肝和脾中MSC与CIK/NK细胞平均比例之和均高于肾。结论是，如果MSC和CIK/NK细胞以相同方式同时注入，它们可能会相互作用，MSC对CIK/NK细胞的抑制在体内发生的部位可能是肺和肝的网状内皮系统。
