Insights into Caco-2 cell culture structure using coherent anti-Stokes Raman scattering (CARS) microscopy

Forskningsoutput: TidskriftsbidragArtikelVetenskapligPeer review

Sammanfattning

We have used coherent anti-Stokes Raman scattering (CARS) microscopy as a novel and rapid, label-free and non-destructive imaging method to gain structural insights into live intestinal epithelial cell cultures used for drug permeability testing. Specifically we have imaged live Caco-2 cells in (bio)pharmaceutically relevant conditions grown on membrane inserts. Imaging conditions were optimized, including evaluation of suitable membrane materials and media solutions, as well as tolerable laser powers for non-destructive imaging of the live cells. Lipid structures, in particular lipid droplets, were imaged within the cells on the insert membranes. The size of the individual lipid droplets increased substantially over the 21-day culturing period up to approximately 10% of the volume of the cross section of individual cells. Variation in lipid content has important implications for intestinal drug permeation testing during drug development but has received limited attention to date due to a lack of suitable analytical techniques. CARS microscopy was shown to be well suited for such analysis with the potential for in situ imaging of the same individual cell-cultures that are used for permeation studies. Overall, the method may be used to provide important information about cell monolayer structure to better understand drug permeation results.
Originalspråkengelska
TidskriftInternational Journal of Pharmaceutics
Volym523
Utgåva1
Sidor (från-till)270–280
Antal sidor11
ISSN0378-5173
DOI
StatusPublicerad - 15 maj 2017
MoE-publikationstypA1 Tidskriftsartikel-refererad

Vetenskapsgrenar

  • 317 Farmaci

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title = "Insights into Caco-2 cell culture structure using coherent anti-Stokes Raman scattering (CARS) microscopy",
abstract = "We have used coherent anti-Stokes Raman scattering (CARS) microscopy as a novel and rapid, label-free and non-destructive imaging method to gain structural insights into live intestinal epithelial cell cultures used for drug permeability testing. Specifically we have imaged live Caco-2 cells in (bio)pharmaceutically relevant conditions grown on membrane inserts. Imaging conditions were optimized, including evaluation of suitable membrane materials and media solutions, as well as tolerable laser powers for non-destructive imaging of the live cells. Lipid structures, in particular lipid droplets, were imaged within the cells on the insert membranes. The size of the individual lipid droplets increased substantially over the 21-day culturing period up to approximately 10{\%} of the volume of the cross section of individual cells. Variation in lipid content has important implications for intestinal drug permeation testing during drug development but has received limited attention to date due to a lack of suitable analytical techniques. CARS microscopy was shown to be well suited for such analysis with the potential for in situ imaging of the same individual cell-cultures that are used for permeation studies. Overall, the method may be used to provide important information about cell monolayer structure to better understand drug permeation results.",
keywords = "317 Pharmacy, Caco-2 cells , Live cell imaging , CARS microscopy , Lipid droplets , Non-linear imaging , Raman spectroscopy , 2-PHOTON FLUORESCENCE MICROSCOPY , LIPID DROPLETS , DRUG ABSORPTION , MONOLAYERS , TRANSPORT , PHOTODAMAGE , DELIVERY , BIOLOGY , NANOPARTICLES , BIOGENESIS",
author = "Jukka Saarinen and Erkan S{\~o}zeri and Sara Fraser-Miller and Leena Peltonen and {A. Santos}, Helder and Antti Isom{\"a}ki and Strachan, {Clare J.}",
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language = "English",
volume = "523",
pages = "270–280",
journal = "International Journal of Pharmaceutics",
issn = "0378-5173",
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Insights into Caco-2 cell culture structure using coherent anti-Stokes Raman scattering (CARS) microscopy. / Saarinen, Jukka ; Sõzeri, Erkan; Fraser-Miller, Sara ; Peltonen, Leena ; A. Santos, Helder; Isomäki, Antti; Strachan, Clare J.

I: International Journal of Pharmaceutics, Vol. 523, Nr. 1, 15.05.2017, s. 270–280.

Forskningsoutput: TidskriftsbidragArtikelVetenskapligPeer review

TY - JOUR

T1 - Insights into Caco-2 cell culture structure using coherent anti-Stokes Raman scattering (CARS) microscopy

AU - Saarinen, Jukka

AU - Sõzeri, Erkan

AU - Fraser-Miller, Sara

AU - Peltonen, Leena

AU - A. Santos, Helder

AU - Isomäki, Antti

AU - Strachan, Clare J.

PY - 2017/5/15

Y1 - 2017/5/15

N2 - We have used coherent anti-Stokes Raman scattering (CARS) microscopy as a novel and rapid, label-free and non-destructive imaging method to gain structural insights into live intestinal epithelial cell cultures used for drug permeability testing. Specifically we have imaged live Caco-2 cells in (bio)pharmaceutically relevant conditions grown on membrane inserts. Imaging conditions were optimized, including evaluation of suitable membrane materials and media solutions, as well as tolerable laser powers for non-destructive imaging of the live cells. Lipid structures, in particular lipid droplets, were imaged within the cells on the insert membranes. The size of the individual lipid droplets increased substantially over the 21-day culturing period up to approximately 10% of the volume of the cross section of individual cells. Variation in lipid content has important implications for intestinal drug permeation testing during drug development but has received limited attention to date due to a lack of suitable analytical techniques. CARS microscopy was shown to be well suited for such analysis with the potential for in situ imaging of the same individual cell-cultures that are used for permeation studies. Overall, the method may be used to provide important information about cell monolayer structure to better understand drug permeation results.

AB - We have used coherent anti-Stokes Raman scattering (CARS) microscopy as a novel and rapid, label-free and non-destructive imaging method to gain structural insights into live intestinal epithelial cell cultures used for drug permeability testing. Specifically we have imaged live Caco-2 cells in (bio)pharmaceutically relevant conditions grown on membrane inserts. Imaging conditions were optimized, including evaluation of suitable membrane materials and media solutions, as well as tolerable laser powers for non-destructive imaging of the live cells. Lipid structures, in particular lipid droplets, were imaged within the cells on the insert membranes. The size of the individual lipid droplets increased substantially over the 21-day culturing period up to approximately 10% of the volume of the cross section of individual cells. Variation in lipid content has important implications for intestinal drug permeation testing during drug development but has received limited attention to date due to a lack of suitable analytical techniques. CARS microscopy was shown to be well suited for such analysis with the potential for in situ imaging of the same individual cell-cultures that are used for permeation studies. Overall, the method may be used to provide important information about cell monolayer structure to better understand drug permeation results.

KW - 317 Pharmacy

KW - Caco-2 cells

KW - Live cell imaging

KW - CARS microscopy

KW - Lipid droplets

KW - Non-linear imaging

KW - Raman spectroscopy

KW - 2-PHOTON FLUORESCENCE MICROSCOPY

KW - LIPID DROPLETS

KW - DRUG ABSORPTION

KW - MONOLAYERS

KW - TRANSPORT

KW - PHOTODAMAGE

KW - DELIVERY

KW - BIOLOGY

KW - NANOPARTICLES

KW - BIOGENESIS

UR - https://www.journals.elsevier.com/international-journal-of-pharmaceutics/

U2 - 10.1016/j.ijpharm.2017.03.015

DO - 10.1016/j.ijpharm.2017.03.015

M3 - Article

VL - 523

SP - 270

EP - 280

JO - International Journal of Pharmaceutics

JF - International Journal of Pharmaceutics

SN - 0378-5173

IS - 1

ER -