Noninvasive estimation of cell cycle phase and proliferation rate of human mesenchymal stem cells by phase-shifting laser microscopy

Tokumitsu, Ayako; Wakitani, Shigeyuki; Takagi, Mutsumi

doi:10.1007/s10616-009-9209-9

Cited by 9 publications

(2 citation statements)

References 9 publications

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“…5B). Thus, it was possible to estimate noninvasively the proliferation activity of MSCs by measuring the phase shift by PLM [18].…”

Section: Estimation Of Cell Cycle and Proliferation Activity By Plmmentioning

confidence: 99%

Noninvasive quality estimation of adherent mammalian cells for transplantation

Takagi

2010

Biotechnol Bioproc E

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The noninvasive quality estimation of adherent mammalian cells for transplantation is reviewed. The quality and heterogeneity of cells should be estimated before transplantation because cultured cells are not homogeneous but heterogeneous. The estimation of cell quality should be performed noninvasively because most protocols of regenerative medicine are autologous cell system. The differentiation level and contamination of other cell lineage could be estimated by two-dimensional cell morphology analysis and tracking using a conventional phase contrast microscope. The noninvasive determination of the laser phase shift of a cell using a phase-shifting laser microscope, which might be more noninvasive, and more useful than the atomic force microscope and digital holographic microscope, was carried out to determine the three-dimensional cell morphology, and the estimation of the cell cycle phase of each adhesive cell and the mean proliferation activity of a cell population. Chemical analysis of the culture supernatant by conventional analytical methods such as ELISA was also useful to estimate the differentiation level of a cell population. Chemical analysis of cell membrane and intracellular components using a probe beam, an infrared beam, and Raman spectroscopy was useful for diagnosing the viability, apoptosis, and differentiation of each adhesive cell. © KSBB hÉóïçêÇëW=ÅÉää=éêçÅÉëëáåÖI=åçåáåî~ëáîÉI=èì~äáíó=ÅçåíêçäI=ãáÅêçëÅçéÉI=ä~ëÉêI=o~ã~å= = = = = CELL PROCESSING ENGINEERING AND CELL QUALITY ESTIMATIONCultivation engineering was established by many researchers aiming at the industrialization of the production of useful substances such as amino acids, antibiotics, and nucleic acids with the use of microbes. The research included medium design, reactor design, flask design, oxygen transfer theory, sensor design, modeling of microbial reactions, and optimization of microbial processes. Cultivation engineering for animal cells was developed from 1980 in order to industrialize the production of animal cell products such as interferon, interleukins, cytokines, tissue plasminogen activator, and therapeutic antibodies, which included the development of serum-free media, new aeration systems without sparging, and the designs of micro carriers for adhesion-dependent cells, reactors, and impellers design. The widespread use and industrialization of regenerative medicine have been highly

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“…5B). Thus, it was possible to estimate noninvasively the proliferation activity of MSCs by measuring the phase shift by PLM [18].…”

Section: Estimation Of Cell Cycle and Proliferation Activity By Plmmentioning

confidence: 99%

Noninvasive quality estimation of adherent mammalian cells for transplantation

Takagi

2010

Biotechnol Bioproc E

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“…Two-dimensional analysis employed a conventional inverted microscope for determining whether the differentiation of hMSCs into chondrocytes had occurred . Threedimensional analysis employed a phase-shifting laser microscope (PLM) to estimate the cell cycle, mean generation time and tumorigenesis (Ito and Takagi 2009;Tokumitsu et al 2009Tokumitsu et al , 2010.…”

Section: Introductionmentioning

confidence: 99%

Effect of the distance between adherent mesenchymal stem cell and the focus of irradiation of femtosecond laser on cell replication capacity

et al. 2012

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The effect of femtosecond laser irradiation on adherent mesenchymal stem cell was investigated with the aim to develop a novel noninvasive cell purification system. A single mesenchymal stem cell was irradiated with a femtosecond laser on the center of the cell using several energy values and the cell lost its replication capacity with one time irradiation with an energy of 2.0 μJ. Besides at a neighbor point in the major axis, when irradiated in the minor axis at a distance shorter than 10 μm, the cell stopped its replication capacity. The accumulation effect of cell damage caused by multiple laser shots at a neighbor point in the minor axis was correlated with the critical distance at which the cell lost its replication capacity. Finally, a novel equation of laser cell damage as a function of laser pulse energy and laser shot number is proposed.

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Cell Processing Engineering for Regenerative Medicine

Takagi

2014

Bioreactor Engineering Research and Industrial Applications II

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The cell processing engineering including automatic cell processing and noninvasive cell quality estimation of adherent mammalian cells for regenerative medicine was reviewed. Automatic cell processing necessary for the industrialization of regenerative medicine was introduced. The cell quality such as cell heterogeneity should be noninvasively estimated before transplantation to patient, because cultured cells are usually not homogeneous but heterogeneous and most protocols of regenerative medicine are autologous system. The differentiation level could be estimated by two-dimensional cell morphology analysis using a conventional phase-contrast microscope. The phase-shifting laser microscope (PLM) could determine laser phase shift at all pixel in a view, which is caused by the transmitted laser through cell, and might be more noninvasive and more useful than the atomic force microscope and digital holographic microscope. The noninvasive determination of the laser phase shift of a cell using a PLM was carried out to determine the three-dimensional cell morphology and estimate the cell cycle phase of each adhesive cell and the mean proliferation activity of a cell population. The noninvasive discrimination of cancer cells from normal cells by measuring the phase shift was performed based on the difference in cytoskeleton density. Chemical analysis of the culture supernatant was also useful to estimate the differentiation level of a cell population. A probe beam, an infrared beam, and Raman spectroscopy are useful for diagnosing the viability, apoptosis, and differentiation of each adhesive cell.

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Noninvasive estimation of cell cycle phase and proliferation rate of human mesenchymal stem cells by phase-shifting laser microscopy

Cited by 9 publications

References 9 publications

Noninvasive quality estimation of adherent mammalian cells for transplantation

Noninvasive quality estimation of adherent mammalian cells for transplantation

Effect of the distance between adherent mesenchymal stem cell and the focus of irradiation of femtosecond laser on cell replication capacity

Cell Processing Engineering for Regenerative Medicine

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