CONCLUSION

Although its influence on the viability and growth of BMSCs needs further elucidation, radiolabeling with ¹¹¹In could be useful for tracking intravenously injected BMSCs in an animal model of brain disease. We observed the dose-dependent growth inhibition of BMSCs by ¹¹¹In-labeling, which was developed by dose-dependent, transient cell cycle arrest, not by cellular senescence or apoptosis/necrosis.

REFERENCES

1. Aguilar S, Nye E, Chan J, Loebinger M, Spencer-Dene B, Fisk N, et al: Murine but not human mesenchymal stem cells generate osteosarcoma like lesions in the lung. Stem Cells 25:1586–1594, 2007

2. Bang OY, Lee JS, Lee PH, Lee G: Autologous mesenchymal stem cell transplantation in stroke patients. Ann Neurol 57: 874-882, 2005

3. Barbash IM, Chouraqui P, Baron J, Feinberg MS, Etzion S, Tessone A, et al: Systemic delivery of bone marrow-derived mesenchymal stem cells to the infarcted myocardium:

feasibility, cell migration, and body distribution. Circulation 108: 863–868, 2003

4. Berger MM, See DM, Redl B, Aymard M, Bruno L: Direct in situ transcriptase polymerase chain reaction for the detection of Enterovirus genome in liver tissues. J Virol Methods 65: 55–66, 1997

5. Bindslev L, Haack-Sorensen M, Bisgaard K, Kragh L, Mortensen S, Hesse B, et al:

Labelling of human mesenchymal stem cells with indium-111 for SPECT imaging: effect on cell proliferation and differentiation. Eur J Nucl Med Mol Imaging 33: 1171–1177, 2006

6. Brenner W, Aicher A, Eckey T, Massoudi S, Zuhayra M, Koehl U, et al: ¹¹¹In-labeled CD34+ hematopoietic progenitor cells in a rat myocardial infarction model. J Nucl Med 45: 512–518, 2004

7. Chen S, Pickard JD, Harris NG: Time course of cellular pathology after controlled cortical impact injury. Exp Neurol 182: 87-102, 2003

8. Chin BB, Nakamoto Y, Bulte JW, Pittenger MF, Wahl R, Kraitchman DL: ¹¹¹In-oxine labelled mesenchymal stem cell SPECT after intravenous administration in myocardial infarction. Nucl Med Commun 24: 1149–1154, 2003

9. Choi Bu-Young, Kim Hak-Yup, Lee Ki-Ho, Cho Youl-Hee, Kong Gu: Clofilium, a potassium channel blocker, induces apoptosis of human promyelocytic leukemia (HL-60) cells via Bcl-2-insensitive activation of caspase-3. Cancer Letters 147: 85-93, 1999

10. Friedenstein AJ, Deriglasova UF, Kulagina NN, Panasuk AF, Rudakowa SF, Luria EA, et al: Precursors for fibroblasts in different populations of hematopoietic cells as detected by the in vitro colony assay method. Exp Hematol 2: 83–92, 1974

11. George J, Kuboki Y, Miyata T: Differentiation of mesenchymal stem cells into osteoblasts on honeycomb collagen scaffolds. Biotechnol Bioeng 95: 404–411, 2006

12. Gunter KP, Lukens JN, Clanton JA, Morris PJ, Janco RL, English D: Neutrophil labeling with indium-111: tropolone vs. oxine. Radiology 149: 563–566, 1983

13. Hofmann M, Wollert KC, Meyer GP, Menke A, Arseniev L, Hertenstein B, et al:

Monitoring of bone marrow cell homing into the infarcted human myocardium.

Circulation 111: 2198–2202, 2005

14. Honma T, Honmou O, Iihoshi S, Harada K, Houkin K, Hamada H, et al: Intravenous infusion of immortalized human mesenchymal stem cells protects against injury in a cerebral ischemia model in adult rat. Exp Neurol 199: 56–66, 2006

15. Huang J, Lee CC, Sutcliffe JL, Cherry SR, Tarantal AF: Radiolabeling rhesus monkey CD34+ hematopoietic and mesenchymal stem cells with ⁶⁴ Cu-pyruvaldehyde-bis(N4-methylthiosemicarbazone) for micro PET imaging. Mol Imaging 7:1-11, 2008

16. Istvan Vermes, Clemens Haanen , Helga Steffens-Nakken , Chris Reutelingsperger: A novel assay for apoptosis Flow cytometric detection of phosphatidylserine early apoptotic cells using fluorescein labeled expression on Annexin V. J Immuno Methods 184: 39-51, 1995

17. Jin Y, Kong H, Stodilka RZ, Wells RG, Zabel P, Merrifield PA, et al: Determining the minimum number of detectable cardiac-transplanted ¹¹¹In-tropolone-labelled

bone-marrow-derived mesenchymal stem cells by SPECT. Phys Med Biol 50: 4445–4455, 2005

18. Kang WJ, Kang HJ, Kim HS, Chung JK, Lee MC, Lee DS: Tissue distribution of 18F-FDG-labeled peripheral hematopoietic stem cells after intracoronary administration in patients with myocardial infarction. J Nucl Med 47: 1295–1301, 2006

19. Karp JM, Leng Teo GS: Mesenchymal stem cell homing: the devil is in the details. Cell Stem Cell 4: 206–216, 2009

20. Kazuaki Mishima, James T. Handa, Amy Aotaki-Keen, Gerard A. Lutty, Lawrence S.

Morse, Leonard M. Hjelmeland: Senescence-Associated β-Galactosidase Histochemistry for the Primate Eye. IOVS, 40: 1590-1593, 1999

21. Kim H, Kim S, Ahn Y: Animal model of traumatic brain injury induced by controlled cortical impact device. J Kor Neurotraumatol Soc 3: 91–98, 2007

22. Kraitchman DL, Tatsumi M, Gilson WD, Ishimori T, Kedziorek D, Walczak P, et al:

Dynamic imaging of allogeneic mesenchymal stem cells trafficking to myocardial infarction. Circulation 112: 1451–1461, 2005

23. Lappalainen RS, Narkilahti S, Huhtala T, Liimatainen T, Suuronen T, Narvanen A, et al:

The SPECT imaging shows the accumulation of neural progenitor cells into internal organs after systemic administration in middle cerebral artery occlusion rats. Neurosci Lett 440: 246–250, 2008

24. Lee-MacAry AE, Ross EL, Davies D, Laylor R, Honeychurch J, Glennie MJ, et al:

Development of a novel flow cytometric cellmediated cytotoxicity assay using the fluorophores PKH-26 and TOPRO-3 iodide. J Immunol Methods 252: 83–92, 2001

25. Lee PH, Kim JW, Bang OY, Ahn YH, Joo IS, Huh K: Autologous mesenchymal stem cell therapy delays the progression of neurological deficits in patients with multiple system atrophy. Clin Pharmacol Ther 83: 723–730, 2008

26. Li Y, Chen J, Chen XG, Wang L, Gautam SC, Xu YX, et al: Human marrow stromal cell therapy for stroke in rat; neurotrophins and functional recovery. Neurology 59: 514–23, 2002

27. Mackay AM, Beck SC, Murphy JM, Barry FP, Chichester CO, Pittenger MF:

Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow.

Tissue Eng 4: 415–428, 1998

28. Min JJ, Ahn Y, Moon S, Kim YS, Park JE, Kim SM, et al: In vivo bioluminescence imaging of cord blood derived mesenchymal stem cell transplantation into rat myocardium. Ann Nucl Med 20: 165–170, 2006

29. Park HJ, Lee PH, Bang OY, Lee G, Ahn YH: Mesenchymal stem cells therapy exerts neuroprotection in a progressive animal model of Parkinson's disease. J Neurochem 107: 141–151, 2008

30. Philip M. Bak and Ralph J. Panos: Protease Antigen Recovery Decreases the Specificity of Bromodeoxyuridine Detection In Formalin-fixed Tissue. J Histochem Cytochem 45:

1165–1170, 1997

31. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al:

Multilineage potential of adult human mesenchymal stem cells. Science : 284:143–

147,1999

32. Riggi N, Cironi L, Provero P, Suva ML, Kaloulis K, Garcia-Echeverria C, et al:

Development of Ewing's sarcoma from primary bone marrowderived mesenchymal progenitor cells. Cancer Res 65:11459–11468, 2005

33. Scheffel U, Tsan MF, Mitchell TG, Camargo EE, Braine H, Ezekowitz MD, et al: Human platelets labeled with In-111 8-hydroxyquinoline: kinetics, distribution, and estimates of

radiation dose. J Nucl Med 23: 149–156, 1982

34. Sekiya I, Larson BL, Vuoristo JT, Cui JG, Prockop DJ: Adipogenic differentiation of human adult stem cells from bone marrow stroma (MSCs). J Bone Miner Res 19: 256–

264, 2004

35. Schonmeyr BH, Wong AK, Soares M, Fernandez J, Clavin N, Mehrara BJ: Ionizing radiation of mesenchymal stem cells results in diminution of the precursor pool and limits potential for multilineage differentiation. Plast Reconstr Surg 222: 64-76, 2008.

.

36. Studeny M, Marini FC, Dembinski JL, Zompetta C, Cabreira-Hansen M, Bekele BN, et al: Mesenchymal stem cells: potential precursors for tumor stroma and targeted-delivery vehicles for anticancer agents. J Natl Cancer Inst 96: 1593–1603, 2004

37. Thakur ML, Coleman RE, Welch MJ: Indium-111-labeled leukocytes for the localization of abscesses: preparation, analysis, tissue distribution, and comparison with gallium-67 citrate in dogs. J Lab Clin Med 89: 217–228, 1977

38. Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C: A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods 184: 39-51, 1995

39. Walczak P, Zhang J, Gilad AA, Kedziorek DA, Ruiz-Cabello J, Young RG, et al: Dual-modality monitoring of targeted intraarterial delivery of mesenchymal stem cells after transient ischemia. Stroke 39: 1569–1574, 2008

40. Woodbury D, Schwarz EJ, Prockop DJ, Black IB: Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res 61: 364–370, 2000

41. Zhu W, Xu W, Jiang R, Qian H, Chen M, Hu J, et al: Mesenchymal stem cells derived from bone marrow favor tumor cell growth in vivo. Exp Mol Pathol 80: 267–274, 2006

- 국문요약 - 5-bromo-4-chloro-3-indolyl-D-galactopyranoside (X-gal)로 염색 후에 현미경으로 분석하였다. Annexin V- fluorescein isothiocyanate (FITC)/ propidium iodide (PI)으로 염색한 후에 FACS 분석을 실시했는데, 높은 방사선량 (11.1 and 33.3 Bq/cell)의 In-111로 표지하여 세포 자멸사나 괴사 변화를 확인하기 위해 초기 (표지 3, 12 시간

후)와 후기 (표지 7 일 후) 단계의 변화를 비교하였다. ¹¹¹In-tropolone으로 표지한

현저하게 높게 관찰되었다. Confocal 현미경 상에서 방사성 표지된 중간엽 줄기 세포가 뇌 질환이 있는 곳에서 확인되었다. In-111 표지 후에, 방사선량에 의존한 중간엽 줄기 세포의 성장이 억제되는 것을 확인했는데, 이는 방사선량 혹은 일시적인 세포 주기 정지에 의한 것이며, 세포성 노화나, 세포 자멸사/괴사에

의한 것은 아닌 것으로 관찰되었다. 비록 ¹¹¹In 표지로 인해 성장 억제가 임상

사용에 앞서 좀 더 검토가 필요하지만, ¹¹¹In-중간엽 줄기 세포는 뇌 질환

동물모델에서 정맥 내 투여한 줄기세포를 추적하는 데는 유용할 것으로 기대한다.

핵심어: ¹¹¹In-tropolone, Bone marrow mesenchymal stem cells, Cell tracking, Growth arrest, Traumatic brain, Radiotoxicity