Real‐time, noninvasive optical coherence tomography of cross‐sectional living cell‐sheets <i>in vitro</i> and <i>in vivo</i>

Kobayashi, M.; Haraguchi, Yuji; Mizuuchi, Kiminori; Iseki, Hiroshi

doi:10.1002/jbm.b.33309

Cited by 8 publications

(20 citation statements)

References 47 publications

(40 reference statements)

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“…The change of cell morphology before and after detachment is thought to be caused by the cytoskeletal tensile reorganization of cells after losing their anchorage between the cells and the culture surface. It has been reported that a spontaneously detached cell sheet contains several cell layers and that the thickness also increases after detaching . The OCM observations were consistent with our previous results.…”

Section: Resultssupporting

confidence: 93%

“…In addition, high‐resolution cross‐sectional images of cells were captured by OCM at a thickness of approximately 10 μm [Figure (C)]. Clear images of cells cultured two‐dimensionally (2‐D) on a culture dish cannot be easily acquired by an OCT system because of its lower resolution (10–20 μm) . On the other hand, OCM could clearly detect the cells because of its higher resolution.…”

Section: Resultsmentioning

confidence: 99%

“…In the system, the light scattering from tissue samples can be detected as a signal‐strength, and the strength reflects a monochrome cross‐sectional image with shades. Recently, we have reported an OCT system, which can observe and analyze the behavior of bioengineered tissues both in vitro and in vivo . The OCT system can acquire a tomographic image of a vertical slice in real‐time and analyze a specific area with a volume of 14 × 14 × 2 mm at a resolution (horizontal: approximately 20 μm; vertical: approximately 9 μm) within 10 s. While the magnification of an OCT image is fixed, that of a microscope is variable by changing the lenses.…”

Section: Introductionmentioning

confidence: 99%

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Optical coherence microscopy of living cells and bioengineered tissue dynamics in high-resolution cross-section

Hasegawa

Haraguchi

Oikaze

et al. 2015

J. Biomed. Mater. Res.

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Optical coherence tomography (OCT) is a valuable tool in the cross-sectional observation/analysis of three-dimensional (3-D) biological tissues, and that histological observation is important clinically. However, the resolution of the technology is approximately 10-20 μm. In this study, optical coherence microscopy (OCM), a tomographic system combining OCT technology with a microscopic technique, was constructed for observing cells individually with a resolution at the submicrometer level. Cells and 3-D tissues fabricated by cell sheet technology were observed by OCM. Importantly, the cell nuclei and cytoplasm could be clearly distinguished, and the time-dependent dynamics of cell-sheet tissues could be observed in detail. Additionally, the 3-D migration of cells in the bioengineered tissue was also detected using OCM and metal-labeled cells. Bovine aortic endothelial cells, but not NIH3T3 murine embryonic skin fibroblasts, actively migrated within the 3-D tissues. This study showed that the OCM system would be a valuable tool in the fields of cell biology, tissue engineering, and regenerative medicine. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 481-488, 2017.

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Section: Resultssupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Optical coherence microscopy of living cells and bioengineered tissue dynamics in high-resolution cross-section

Hasegawa

Haraguchi

Oikaze

et al. 2015

J. Biomed. Mater. Res.

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“…(C/CSS): OCT analysis of living cell sheets is an important tool in the field of cell sheet‐based TERM to analyze the inner structure of layered living cells without invasion or damage. OCT observations has been used to analyze the static and dynamic behaviors of living cell sheets in vitro and in vivo including cell‐sheet adhesion and the real‐time propagation of cells, demonstrating that it is a powerful tool in a real‐time, non‐invasive manner (Figure ) . OCT has been widely used to investigate the longitudinal development of engineered tissues and cell dynamics such as migration, proliferation, detachment, and cell‐material interactions .…”

Section: Available Imaging Techniquesmentioning

confidence: 99%

Three‐dimensional imaging technologies: a priority for the advancement of tissue engineering and a challenge for the imaging community

Teodori

Crupi

Costa

et al. 2016

Journal of Biophotonics

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Tissue engineering/regenerative medicine (TERM) is an interdisciplinary field that applies the principle of engineering and life sciences to restore/replace damaged tissues/organs with in vitro artificially-created ones. Research on TERM quickly moves forward. Today newest technologies and discoveries, such as 3D-/bio-printing, allow in vitro fabrication of ex-novo made tissues/organs, opening the door to wide and probably never-ending application possibilities, from organ transplant to drug discovery, high content screening and replacement of laboratory animals. Imaging techniques are fundamental tools for the characterization of tissue engineering (TE) products at any stage, from biomaterial/scaffold to construct/organ analysis. Indeed, tissue engineers need versatile imaging methods capable of monitoring not only morphological but also functional and molecular features, allowing three-dimensional (3D) and time-lapse in vivo analysis, in a non-destructive, quantitative, multidimensional analysis of TE constructs, to analyze their pre-implantation quality assessment and their fate after implantation. This review focuses on the newest developments in imaging technologies and applications in the context of requirements of the different steps of the TERM field, describing strengths and weaknesses of the current imaging approaches.

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“…OCT is already used clinically in ophthalmology and the other fields of medical care to analyze cross‐sections of target regions . Using the OCT system, images of cross‐sections of living cell sheets have been obtained …”

Section: Introductionmentioning

confidence: 99%

Real‐time quantitation of internal metabolic activity of three‐dimensional engineered tissues using an oxygen microelectrode and optical coherence tomography

2016

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Recent progress in tissue engineering technology has enabled us to develop thick tissue constructs that can then be transplanted in regenerative therapies. In clinical situations, it is vital that the engineered tissues to be implanted are safe and functional before use. However, there is currently a limited number of studies on real-time quality evaluation of thick living tissue constructs. Here we developed a system for quantifying the internal activities of engineered tissues, from which we can evaluate its quality in real-time. The evaluation was achieved by measuring oxygen concentration profiles made along the vertical axis and the thickness of the tissues estimated from cross-sectional images obtained noninvasively by an optical coherence tomography system. Using our novel system, we obtained (i) oxygen concentration just above the tissues, (ii) gradient of oxygen along vertical axis formed above the tissues within culture medium, and (iii) gradient of oxygen formed within the tissues in real-time. Investigating whether these three parameters could be used to evaluate engineered tissues during culturing, we found that only the third parameter was a good candidate. This implies that the activity of living engineered tissues can be monitored in real-time by measuring the oxygen gradient within the tissues. The proposed measuring strategy can be applied to developing more efficient culturing methods to support the fabrication of engineered thick tissues, as well as providing methods to confirm the quality in real-time. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 855-864, 2017.

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Real‐time, noninvasive optical coherence tomography of cross‐sectional living cell‐sheets in vitro and in vivo

Cited by 8 publications

References 47 publications

Optical coherence microscopy of living cells and bioengineered tissue dynamics in high-resolution cross-section

Optical coherence microscopy of living cells and bioengineered tissue dynamics in high-resolution cross-section

Three‐dimensional imaging technologies: a priority for the advancement of tissue engineering and a challenge for the imaging community

Real‐time quantitation of internal metabolic activity of three‐dimensional engineered tissues using an oxygen microelectrode and optical coherence tomography

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