High-Dose 111 In Induces G1 Cell Cycle Arrest and Cell Death in Rat Bone Marrow Mesenchymal Stem Cells

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ORIGINAL ARTICLE

High-Dose 111In Induces G1 Cell Cycle Arrest and Cell Death in Rat Bone Marrow Mesenchymal Stem Cells Bok-Nam Park & Wooyoung Shim & Young Hwan Ahn & Jae-Ho Lee & Young-Sil An & Joon-Kee Yoon

Received: 14 October 2011 / Revised: 28 November 2011 / Accepted: 14 December 2011 / Published online: 14 January 2012 # Korean Society of Nuclear Medicine 2012

Abstract Purpose This study was performed to evaluate the effect of 111 In-labeling on the cell growth, cycle and viability of bone marrow mesenchymal stem cells (BMSCs). Methods Rat BMSCs were labeled with various doses of 111 In (0.4–11.1 Bq/cell). The growth curve of 111In-BMSCs was obtained up to 14th day of labeling. The cell cycle was evaluated by 5-bromo-2-deoxyuridine (BrdU) labeling or propidium iodide (PI) staining. Senescent cells were counted under a light microscope after staining with 5bromo-4-chloro-3-indolyl- D -galactopyranoside. Flow cytometry was performed to measure apoptotic and necrotic fractions after staining with annexin V-FITC and PI. B.-N. Park : Y.-S. An : J.-K. Yoon (*) Department of Nuclear Medicine and Molecular Imaging, Ajou University School of Medicine, Suwon 442-749, Republic of Korea e-mail: [email protected] W. Shim : Y. H. Ahn Institute for Neuroregeneration and Stem Cell Research, Ajou University School of Medicine, Suwon, Republic of Korea W. Shim Department of Molecular Science and Technology, Ajou University, Suwon, Republic of Korea Y. H. Ahn Department of Neurosurgery, Ajou University School of Medicine, Suwon, Republic of Korea J.-H. Lee Department of Biochemistry, Ajou University School of Medicine, Suwon, Republic of Korea

Results The growth of BMSCs labeled with higher doses of In (4.4 or 11.1 Bq/cell) was significantly inhibited from the 3rd day of labeling. Flow cytometry revealed less BrdUpositive BMSCs at 11.1 Bq 111In/cell during all measurement days and G1 arrest at 4.4 and 11.1 Bq 111In/cell. Significant increases in apoptosis and necrosis were also observed at 4.4 (3.04%/1.35%) and 11.1 Bq 111In/cell (9.07%/3.18%) on the 14th day (control01.60%/0.39%). However, no cellular senescence was visualized up to the 14th day. Conclusion A high dose of 111In-labeling induced cell cycle arrest and death in BMSCs; therefore, it should be used with a careful dosimetry in case of applying it to humans. 111

Keywords 111In-tropolone . Mesenchymal stem cells . Cell cycle . Apoptosis . Necrosis

Introduction For more than a decade, investigators have sought an effective method to use stem cells as a regenerative therapy for various diseases. Allogeneic as well as autologous bone marrow mesenchymal stem cells (BMSCs) have been attempted to repair heart or brain diseases, which are supported by the fact that adult stem cells raise few ethical issues as opposed to embryo-derived stem cells [1–3]. In those trials, BMSCs improved left ventricular function in addition to symptoms in patients with myocardial infarction and induced functional recovery in patients with strokes [2, 3]. With the success of cell therapies,