Korean Chemical Engineering Research

The Korean Institute of Chemical Engineers (한국화학공학회)
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Atomic Force Microscopy(AFM) based Single Cell Manipulation and High Efficient Gene Delivery Technology

원자간력 현미경을 이용한 단일세포 조작 및 고효율 유전자 도입기술

  • Han, Sung-Woong (Computational Cell Biomechanics Team, VCAD System Research Program) ;
  • Nakamura, Chikashi (Research Institute for Cell Engineering, AIST) ;
  • Miyake, Jun (Research Institute for Cell Engineering, AIST) ;
  • Kim, Woo-Sik (Department of Chemical Engineering, Kyung Hee University) ;
  • Kim, Jong-Min (Department of Chemical Engineering, Dong A University) ;
  • Chang, Sang-Mok (Department of Chemical Engineering, Dong A University)
  • 한성웅 (일본이화학연구소 세포시뮬레이션팀) ;
  • ;
  • ;
  • 김우식 (경희대학교 화학공학과) ;
  • 김종민 (동아대학교 화학공학과) ;
  • 장상목 (동아대학교 화학공학과)
  • Received : 2009.07.08
  • Accepted : 2009.08.03
  • Published : 2009.10.31
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Abstract

The principle and application of a scanning probe microscopy(SPM) are reviewed briefly, and a low-invasive single cell manipulation and a gene delivery technique using an etched atomic force microscopy(AFM) probe tip, which we call a nanoneedle, are explained in detail. The nanoneedle insertion into a cell can be judged by a sudden drop of force in a force-distance curve. The probabilities of nanoneedle insertion into cells were 80~90%, which were higher than those of typical microinjection capillaries. When the diameter of the nanoneedle was smaller than 400 nm, the nanoneedle insertion into a cell over 1 hour had almost no influence on the cell viability. A highly efficient gene delivery and a high ratio of expressed gene per delivered DNA compared the conventional major nonviral gene delivery methods could be achieved using the gene modified nanoneedle.

본 총설에서는 주사형프로브현미경의 원리와 응용에 관하여 간략히 설명하고 최근 본 그룹에 의하여 활발하게 연구되고 있는 나노탐침과 AFM(원자간력현미경 atomic force microscopy)을 이용한 저침습성(low-invasive) 단일세포 조작기술과 고효율 유전자 도입기술을 소개하고자 한다. 시판 AFM 탐침을 침상구조로 가공한 나노탐침과 AFM을 이용하였을 경우, 탐침의 세포삽입의 성공여부를 force-distance curve 상의 척력소실의 유무로 판단할 수 있다. 침상 나노탐침을 사용하면 대부분의 세포에서 80~90%의 고효율 세포삽입이 가능하여 마이크로인젝션용 미세관을 이용하는 경우보다 세포삽입효율이 높았다. 또한 나노탐침의 직경이 400 nm 이하의 경우에는 세포 종류에 관계없이 장시간 나노탐침의 삽입에도 세포활성에 큰 영향이 없었다. 침상나노탐침을 이용하여 DNA를 도입하였을 경우에도 기존의 DNA 도입방법과 비교하여 높은 도입효율과 유전자 발현율로 DNA를 도입할 수 있는 가능성을 확인하였다.

Keywords

Acknowledgement

Supported by : 동아대학교

References

  1. Chang, S. M. and Kim, J. M., 'The Recent Trands of Advanced Scanning Microprobe Microscopy,' Chem. World., 36(5), 21-25(1996)
  2. Chang, S. M., Lee, H. J., Kim, J. M. and Muramatsu, H., 'AFM Mode Scanning Near-field Optical Microscopy(SNOAM) and Nanotechnology,' Prospectives of Ind. Chem., 2(4), 52-60(1999)
  3. Takahashi, K. and Yamanaka, S., "Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors," Cell, 126, 663-676(2006) https://doi.org/10.1016/j.cell.2006.07.024
  4. Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T., Tomoda, K. and Yamanaka, S., "Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors," Cell, 131, 1-12(2007) https://doi.org/10.1016/j.cell.2007.09.034
  5. Hahn, W., Ho, S. H., Jeong, J. G., Hahn, E. Y., Kim, S., Yu, S. S., Kim, S. and Kim, J. M., "Viral Vector-Mediated Transduction of a Modified Thrombospondin-2 cDNA Inhibits Tumor Growth and Angiogenesis," Gene Ther., 11, 739-45(2004) https://doi.org/10.1038/sj.gt.3302219
  6. Selkirk, S. M., Green, S. J., Plunkett, R. J., Baron, T. A., Lis, A. and Spence, P. O., "Syngeneic Central Nervous System Transplantation of Genetically Transduced Mature, Adult Astrocytes," Gene Ther., 9, 432-443(2002) https://doi.org/10.1038/sj.gt.3301643
  7. Kang, Y., Stein, C. S., Heth, J. A., Sinn, P. L., Penisten, A. K., Staber, P. D., Ratliff, K. L., Shen, H., Barker, C. K., Martins, I., Sharkey, C. M., Sanders, D. A., McCray, P. B. and Davidson, B. L., “In Vivo Gene Transfer Using a Nonprimate Lentiviral Vector Pseudotyped with Ross River Virus Glycoproteins,” J. Virol., 76, 9378-9388(2002) https://doi.org/10.1128/JVI.76.18.9378-9388.2002
  8. Modlich, U., Bohne, J., Schmidt, M., von Kalle, C., Knoss, S., Schambach, A. and Baum, C., "Cell-Culture Assays Reveal the Importance of Retroviral Vector Design for Insertional Genotoxicity," Blood, 108, 2545-2553(2006) https://doi.org/10.1182/blood-2005-08-024976
  9. Mathis, J. M., Williams, B. J., Sibley, D. A., Carroll, J. L., Li, J., Odaka, Y., Barlow, S., Nathan, C. O., Li, B. D. and DeBenedetti, A., "Cancer-Specific Targeting of an Adenovirusdelivered Herpes Simplex Virus Thymidine Kinase Suicide Gene Using Translational Control," J. Gene Med., 8, 1105-1120(2006) https://doi.org/10.1002/jgm.935
  10. Bruder, J. T. and Kovesdi, I., 'Adenovirus Infection Stimulates the Raf/MAPK Signaling Pathway and Induces Interleukin-8 Expression,' J. Virol., 71, 398-404(1997)
  11. Shifrin, A. L., Chirmule, N., Gao, G. P., Wilson, J. M. and Raper, S. E., "Innate Immune Responses to Adenoviral Vector-mediated Acute Pancreatitis," Pancreas, 30, 122-129(2005) https://doi.org/10.1097/01.mpa.0000151578.99413.88
  12. Bieber, T., Meissner, W., Kostin, S., Niemann, A. and Elsasser, H. P., "Intracellular Route and Transcriptional Competence of Polyethylenimine-DNA Complexes," J. Control. Release, 82, 441-454(2002) https://doi.org/10.1016/S0168-3659(02)00129-3
  13. Hoelters, J., Ciccarella, M., Drechsel, M., Geissler, C., Gulkan, H., Bocker, W., Schieker, M., Jochum, M. and Neth, P., "Nonviral Genetic Modification Mediates Effective Transgene Expression and Functional RNA Interference in Human Mesenchymal Stem Cells," J. Gene Med., 7, 718-728(2005) https://doi.org/10.1002/jgm.731
  14. Jeschke, M. G., Barrow, R. E., Hawkins, H. K., Yang, K., Hayes, R. L., Lichtenbelt, B. J., Perez-Polo, J. R. and Herndon, D. N., "IGF-I Gene Transfer in Thermally Injured Rats," Gene Ther., 6, 1015-1020(1999) https://doi.org/10.1038/sj.gt.3300923
  15. Pollard, H., Remy, J. S., Loussouarn, G., Demolombe, S., Behr, J. P. and Escande, D., "Polyethylenimine But not Cathionic Lipids Promotes Transgene Delivery to the Nucleus in Mammalian Cells," J. Biol. Chem., 273, 7507-7511(1998) https://doi.org/10.1074/jbc.273.13.7507
  16. Vijayanathan, V., Thomas, T. and Thomas, T. J., "DNA Nanoparticles and Development of DNA Delivery Vehicles for Gene Therapy," Biochemistry, 41(48), 14085-14094(2002) https://doi.org/10.1021/bi0203987
  17. Knoblauch, M., Hibberd, J. M., Gray, J. C. and van Bell, A. J., "A Galinstan Expansion Femtosyringe for Microinjection of Eukaryotic Organelles and Prokaryotes," Nat. Biotechnol., 17, 906- 909(1999) https://doi.org/10.1038/12902
  18. Tsulaia, T. V., Prokopishyn, N. L., Yao, A., Carsrud, N. D. V., Carou, M. C., Brown, D. B., Davis, B. R. and Yannariello- Brown, J., "Glass Needle-mediated Microinjection of Macromolecules and Transgenes Into Primary Human Mesenchymal Stem Cells," J. Biomed. Sci., 10, 328-336(2003) https://doi.org/10.1007/BF02256452
  19. Han, S., Nakamura, C., Imai, Y., Nakamura, N. and Miyake, J., "Monitoring of Hormonal Drug Effect in a Single Breast Cancer Cell Using an Estrogen Responsive GFP Reporter Vector Delivered by a Nanoneedle," Biosens. Bioelectron., 24, 1219-1222(2008) https://doi.org/10.1016/j.bios.2008.07.017
  20. Han, S., Nakamura, C., Kotobuki, N., Obataya, I., Ohgushi, H., Nagamune, T. and Miyake, J., 'High-efficiency DNA Injection Into a Single Human Mesenchymal Stem Cell Using a Nanoneedle and Atomic Force Microscopy', Nanomedicine, 4, 215-225(2008) https://doi.org/10.1016/j.nano.2008.03.005
  21. Han, S., Nakamura, C., Obataya, I., Nakamura, N. and Miyake, J., “A Molecular Delivery System by Using AFM and Nanoneedle,” Biosens. Bioelectron., 20, 2120-2125(2005) https://doi.org/10.1016/j.bios.2004.08.023
  22. Han, S., Nakamura, C., Obataya, I., Nakamura, N. and Miyake, J., "Gene Expression Using an Ultra Thin Needle Enabling Accurate Displacement and Low Invasiveness," Biochem. Bioph. Res. Co., 332, 633-639(2005) https://doi.org/10.1016/j.bbrc.2005.04.059
  23. Nakamura, C., Kamiishi, H., Nakamura, N. and Miyake, J., 'A Nanoneedle Can be Inserted into a Living Cell Without Any Mechanical Stress Inducing Calcium ion Influx,' Electrochemistry, 76, 586-589(2008) https://doi.org/10.5796/electrochemistry.76.586
  24. Obataya, I., Nakamura, C., Han, S., Nakamura, N. and Miyake, J., "Nanoscale Operation of a Living Cell Using An Atomic Force Microscope with a Nanoneedle," Nano Lett., 5, 27-30(2005) https://doi.org/10.1021/nl0485399
  25. Obataya, I., Nakamura, C., Han, S., Nakamura, N. and Miyake, J., "Mechanical Sensing of the Penetration of Various Nanoneedles Into a Living Cell Using Atomic Force Microscopy," Biosens. Bioelectron., 20, 1652-1655(2005) https://doi.org/10.1016/j.bios.2004.07.020
  26. Obataya, I., Nakamura, C., Han, S., Nakamura, N. and Miyake, J., "Direct Insertion of Proteins Into a Living Cell Using Atomic Force Microscope with a Nanoneedle," Nanobiotechnology, 1, 347-351(2005) https://doi.org/10.1385/NBT:1:4:347
  27. Mathur, A. B., Truskey, G. A. and Reichert, W. M., 'Total Internal Reflection Microscopy and Atomic Force Microscopy(TIRFMAFM) to Study Stress Transduction Mechanisms in Endothelial Cells,' Crit. Rev. Biomed. Eng., 28, 197-222(2000) https://doi.org/10.1615/CritRevBiomedEng.v28.i12.340
  28. Rotsch, C., Jacobson, K. and Radmacher, M., "Dimensional and Mechanical Dynamics of Active and Stable Edges in Motile Fibroblasts Investigated by Using Atomic Force Microscopy," Proc. Natl. Acad. Sci., 96, 921-926(1999) https://doi.org/10.1073/pnas.96.3.921
  29. Boron, W. F., "Regulation of Intracellular pH," Adv. Physiol. Educ., 28, 160-179(2004) https://doi.org/10.1152/advan.00045.2004
  30. Doi, A., Mizuno, M., Katafuchi, T., Furue, H., Koga, K. and Yoshimura, M., "Slow Oscillation of Membrane Currents Mediated by Glutamatergic Inputs of Rat Somatosensory Cortical Neurons: In Vivo Patch-clamp Analysis," Eur. J. Neurosci., 26, 2565-2575(2007) https://doi.org/10.1111/j.1460-9568.2007.05885.x