Cheng Kang, Benten Daniel, Bhargava Kuldeep, Inada Mari, Joseph Brigid, Palestro Christopher, Gupta Sanjeev
Marion Bessin Liver Research Center, Diabetes Research Center, and Cancer Research Center, Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Hepatology. 2009 Oct;50(4):1194-203. doi: 10.1002/hep.23120.
Tracking stem/progenitor cells through noninvasive imaging is a helpful means of assessing the targeting of transplanted cells to specific organs. We performed in vitro and in vivo studies wherein adult human hepatocytes and human fetal liver stem/progenitor cells were labeled with indium-111 ((111)In)-oxine and technetium-99m ((99m)Tc)-Ultratag or (99m)Tc-Ceretec. The labeling efficiency and viability of cells was analyzed in vitro, and organ biodistribution of cells was analyzed in vivo after transplantation in xenotolerant nonobese diabetic/severe combined immunodeficiency mice through intrasplenic or intraportal routes. We found that adult hepatocytes and fetal liver stem/progenitor cells incorporated (111)In but not (99m)Tc labels. After radiolabeling, cell viability was unchanged. Transplanted adult hepatocytes or fetal liver stem/progenitor cells were targeted to the liver more effectively by the intraportal rather than the intrasplenic route. Transplanted cells were retained in the liver after intraportal injection and in the liver and spleen after intrasplenic injection, without translocations into pulmonary or systemic circulations. Compared with fetal liver stem/progenitor cells, fewer adult hepatocytes were retained in the spleen after intrasplenic transplantation. The distribution of transplanted cells in organs was substantiated by genetic assays, including polymerase chain reaction amplification of DNA sequences from a primate-specific Charcot-Marie-Tooth element, and in situ hybridization for primate alphoid satellite sequences ubiquitous in all centromeres.
(111)In labeling of human fetal liver stem/progenitor cells and adult hepatocytes was effective for noninvasive localization of transplanted cells. This should facilitate continued development of cell therapies through further animal and clinical studies.
通过非侵入性成像追踪干细胞/祖细胞是评估移植细胞靶向特定器官的一种有用方法。我们进行了体外和体内研究，其中成人肝细胞和人胎肝干细胞/祖细胞用铟 - 111（¹¹¹In）- 奥克辛、锝 - 99m（⁹⁹ᵐTc）- 超标记物或⁹⁹ᵐTc - 脑显像剂进行标记。在体外分析细胞的标记效率和活力，在通过脾内或门静脉途径将细胞移植到具有异种耐受性的非肥胖糖尿病/严重联合免疫缺陷小鼠体内后，在体内分析细胞的器官生物分布。我们发现成人肝细胞和胎肝干细胞/祖细胞摄取了¹¹¹In标记但未摄取⁹⁹ᵐTc标记。放射性标记后，细胞活力未改变。移植的成人肝细胞或胎肝干细胞/祖细胞通过门静脉途径比脾内途径更有效地靶向肝脏。门静脉内注射后，移植细胞保留在肝脏中；脾内注射后，移植细胞保留在肝脏和脾脏中，没有转移到肺循环或体循环中。与胎肝干细胞/祖细胞相比，脾内移植后保留在脾脏中的成人肝细胞较少。通过基因检测证实了移植细胞在器官中的分布，包括对灵长类特异性夏科 - 马里 - 图斯元件的DNA序列进行聚合酶链反应扩增，以及对所有着丝粒中普遍存在的灵长类α卫星序列进行原位杂交。
¹¹¹In标记人胎肝干细胞/祖细胞和成人肝细胞对于移植细胞的非侵入性定位是有效的。这应有助于通过进一步的动物和临床研究继续开展细胞治疗。
