When nerve growth factor (NGF) is interacted with PC12 cells derived from rat pheochromocytoma, they are partially differentiated into neuron-like cells with neurites. In this work, PC12 cells differentiated by NGF were selectively isolated using a localized impulsive force in a μm-scale area, which was generated by focusing an infrared femtosecond laser into a cell culture medium. In order to evaluate the ability of the isolation method, differentiated and undifferentiated cells were isolated and their morphological changes after the isolation were compared. In both cases, their neurites were once contracted and some of them gradually regenerated day by day. When differentiated cells were isolated, the percentage of differentiated cells with regenerated neurites, 6 h after the isolation, was about 3.3 times higher than that when undifferentiated ones were isolated. This result was compared with a control trypsin experiment. In the comparison, it was indicated that the same degree of cell function was maintained when the present isolation method was used.
Femtosecond laser Laser manufacturing Impulsive force Cell Isolation Cell differentiation
This is a preview of subscription content, log in to check access.
This work was partly supported by CREST from JST (Japan Science and Technology Agency) and by the Grant-in-Aid for Young Scientists (A) from JSPS (Japan Society for the Promotion of Science) of Japan to YH. The support by the Grant-Aid on Priority Area “Bio Manipulation,” from MEXT (Ministry of Education, Culture, Sports, Science and Technology) of Japan and by MOE-ATU project (National Chiao Tung University) from the Ministry of Education, Taiwan, and National Science Consul of Taiwan (0970027441) to HM is also acknowledged.
A. Ashkin, J.M. Dziedzic et al., Optical trapping and manipulation of single cells using infrared-laser beams. Nat. 330(6150), 769–771 (1987)CrossRefGoogle Scholar
P.J. Crosland-Taylor, A device for counting small particles suspended in a fluid through a tube. Nat. 171(4340), 37–38 (1953)CrossRefGoogle Scholar
G.C. Easty, D.M. Easty et al., Studies of cellular adhesiveness. Exptl. Cell Res. 19, 539–548 (1960)CrossRefGoogle Scholar
M.J. Evans, M.H. Kaufman, Establishment in culture of pluripotential cells from mouse embryos. Nat. 292(5819), 154–156 (1981)CrossRefGoogle Scholar
M.J. Fluwer, Electronic separation of biological cells by volume. Sci. 150(698), 910–911 (1965)CrossRefGoogle Scholar
L.A. Greene, A.S. Tischler, Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc. Nat. Acad. Sci. USA 73(7), 2424–2428 (1976)CrossRefGoogle Scholar
Y. Hosokawa, H. Takabayashi et al., Nondestructive isolation of single cultured animal cells by femtosecond laser-induced shockwave. Appl. Phys. A 79(4–6), 795–798 (2004)Google Scholar
Y.-S. Huang, S.-N. Cheng et al., Effects of interleukin-15 on neuronal differentiation of neural stem cells. Res. 1304, 38–48 (2009)Google Scholar
H.R. Hulett, W.A. Bonner et al., Cell sorting: automated separation of mammalian cells as a function of intracellular fluorescence. Sci. 166(3906), 747–749 (1969)CrossRefGoogle Scholar
T. Kaji, S. Ito et al., Nondestructive micropatterning of living animal cells using focused femtosecond laser-induced impulsive force. Appl. Phys. Lett. 91(2), 023904 (2007)CrossRefGoogle Scholar
F. Lei, R. Haque et al., T lineage differentiation from induced pluripotent stem cells. Cell. Immunol. 260(1), 1–5 (2009)CrossRefGoogle Scholar
Y. Maezawa, Y. Hosokawa et al., In situ observation of cell-detachment process initiated by femtosecond laser-induced stress wave. Appl. Phys. A. (2010) doi:10.1007/s00339-010-5771-4
G.R. Martin, Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc. Natl. Acad. Sci. USA 78(12), 7634–7638 (1981)CrossRefGoogle Scholar
K. Okita, T. Ichisaka et al., Generation of germline-competent induced pluripotent stem cells. Nat. 448(7151), 313–317 (2007)CrossRefGoogle Scholar
B. Roda, P. Reschiglian et al., Human lymphocyte sorting by gravitational field-flow fractionation. Anal. Bioanal. Chem. 392(1–2), 137–145 (2008)CrossRefGoogle Scholar
K. Takahashi, S. Yamanaka, Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126(4), 663–676 (2006)CrossRefGoogle Scholar
N. Takizawa, K. Okano et al., Viability evaluation of culture cells patterned by femtosecond laser-induced impulsive force. Proc. SPIE 6854, 685411 (2008)CrossRefGoogle Scholar
L. Weiss, Studies on cellular adhesion in tissue-culture: V. Some effects of enzymes on cell-detachment. Exptl. Cell Res. 30(3), 509–520 (1963)CrossRefGoogle Scholar
C.J. Xian, X.F. Zhou, EGF family of growth factors: Essential roles and functional redundancy in the nerve system. Front. Biosci. 9, 85–92 (2004)CrossRefGoogle Scholar
M. Zborowski, L. Sun et al., Rapid cell isolation by magnetic flow sorting for applications in tissue engineering. Asaio J. 45(3), 127–130 (1999)CrossRefGoogle Scholar