Forskning
Udskriv Udskriv
Switch language
Rigshospitalet - en del af Københavns Universitetshospital
Udgivet

Glucose transport by epithelia prepared from harvested enterocytes

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Harvard

Kimura, Y, van der Merwe, M, Bering, SB, Penmatsa, H, Conoley, VG, Sangild, PT, Naren, AP & Buddington, RK 2015, 'Glucose transport by epithelia prepared from harvested enterocytes' Cytotechnology, bind 67, nr. 1, s. 39-49. https://doi.org/10.1007/s10616-013-9656-1

APA

Kimura, Y., van der Merwe, M., Bering, S. B., Penmatsa, H., Conoley, V. G., Sangild, P. T., ... Buddington, R. K. (2015). Glucose transport by epithelia prepared from harvested enterocytes. Cytotechnology, 67(1), 39-49. https://doi.org/10.1007/s10616-013-9656-1

CBE

Kimura Y, van der Merwe M, Bering SB, Penmatsa H, Conoley VG, Sangild PT, Naren AP, Buddington RK. 2015. Glucose transport by epithelia prepared from harvested enterocytes. Cytotechnology. 67(1):39-49. https://doi.org/10.1007/s10616-013-9656-1

MLA

Vancouver

Kimura Y, van der Merwe M, Bering SB, Penmatsa H, Conoley VG, Sangild PT o.a. Glucose transport by epithelia prepared from harvested enterocytes. Cytotechnology. 2015 jan;67(1):39-49. https://doi.org/10.1007/s10616-013-9656-1

Author

Kimura, Yasuhiro ; van der Merwe, Marie ; Bering, Stine B ; Penmatsa, Himabindu ; Conoley, Veronica G ; Sangild, Per T ; Naren, Anjaparavanda P ; Buddington, Randal K. / Glucose transport by epithelia prepared from harvested enterocytes. I: Cytotechnology. 2015 ; Bind 67, Nr. 1. s. 39-49.

Bibtex

@article{5b4b16d9e6e6493cbc9a9f31beada0c9,
title = "Glucose transport by epithelia prepared from harvested enterocytes",
abstract = "Transformed and cultured cell lines have significant shortcomings for investigating the characteristics and responses of native villus enterocytes in situ. Interpretations of results from intact tissues are complicated by the presence of underlying tissues and the crypt compartment. We describe a simple, novel, and reproducible method for preparing functional epithelia using differentiated enterocytes harvested from the small intestine upper villus of adult mice and preterm pigs with and without necrotizing enterocolitis. Concentrative, rheogenic glucose uptake was used as an indicator of epithelial function and was demonstrated by cellular accumulation of tracer (14)C D-glucose and Ussing chamber based short-circuit currents. Assessment of the epithelia by light and immunofluorescent microscopy revealed the harvested enterocytes remain differentiated and establish cell-cell connections to form polarized epithelia with distinct apical and basolateral domains. As with intact tissues, the epithelia exhibit glucose induced short-circuit currents that are increased by exposure to adenosine and adenosine 5'-monophosphate (AMP) and decreased by phloridzin to inhibit the apical glucose transporter SGLT-1. Similarly, accumulation of (14)C D-glucose by the epithelia was inhibited by phloridzin, but not phloretin, and was stimulated by pre-exposure to AMP and adenosine, apparently by a microtubule-based mechanism that is disrupted by nocodazole, with the magnitudes of responses to adenosine, forskolin, and health status exceeding those we have measured using intact tissues. Our findings indicate that epithelia prepared from harvested enterocytes provide an alternative approach for comparative studies of the characteristics of nutrient transport by the upper villus epithelium and the responses to different conditions and stimuli.",
author = "Yasuhiro Kimura and {van der Merwe}, Marie and Bering, {Stine B} and Himabindu Penmatsa and Conoley, {Veronica G} and Sangild, {Per T} and Naren, {Anjaparavanda P} and Buddington, {Randal K}",
year = "2015",
month = "1",
doi = "10.1007/s10616-013-9656-1",
language = "English",
volume = "67",
pages = "39--49",
journal = "Cytotechnology",
issn = "0920-9069",
publisher = "Springer Netherlands",
number = "1",

}

RIS

TY - JOUR

T1 - Glucose transport by epithelia prepared from harvested enterocytes

AU - Kimura, Yasuhiro

AU - van der Merwe, Marie

AU - Bering, Stine B

AU - Penmatsa, Himabindu

AU - Conoley, Veronica G

AU - Sangild, Per T

AU - Naren, Anjaparavanda P

AU - Buddington, Randal K

PY - 2015/1

Y1 - 2015/1

N2 - Transformed and cultured cell lines have significant shortcomings for investigating the characteristics and responses of native villus enterocytes in situ. Interpretations of results from intact tissues are complicated by the presence of underlying tissues and the crypt compartment. We describe a simple, novel, and reproducible method for preparing functional epithelia using differentiated enterocytes harvested from the small intestine upper villus of adult mice and preterm pigs with and without necrotizing enterocolitis. Concentrative, rheogenic glucose uptake was used as an indicator of epithelial function and was demonstrated by cellular accumulation of tracer (14)C D-glucose and Ussing chamber based short-circuit currents. Assessment of the epithelia by light and immunofluorescent microscopy revealed the harvested enterocytes remain differentiated and establish cell-cell connections to form polarized epithelia with distinct apical and basolateral domains. As with intact tissues, the epithelia exhibit glucose induced short-circuit currents that are increased by exposure to adenosine and adenosine 5'-monophosphate (AMP) and decreased by phloridzin to inhibit the apical glucose transporter SGLT-1. Similarly, accumulation of (14)C D-glucose by the epithelia was inhibited by phloridzin, but not phloretin, and was stimulated by pre-exposure to AMP and adenosine, apparently by a microtubule-based mechanism that is disrupted by nocodazole, with the magnitudes of responses to adenosine, forskolin, and health status exceeding those we have measured using intact tissues. Our findings indicate that epithelia prepared from harvested enterocytes provide an alternative approach for comparative studies of the characteristics of nutrient transport by the upper villus epithelium and the responses to different conditions and stimuli.

AB - Transformed and cultured cell lines have significant shortcomings for investigating the characteristics and responses of native villus enterocytes in situ. Interpretations of results from intact tissues are complicated by the presence of underlying tissues and the crypt compartment. We describe a simple, novel, and reproducible method for preparing functional epithelia using differentiated enterocytes harvested from the small intestine upper villus of adult mice and preterm pigs with and without necrotizing enterocolitis. Concentrative, rheogenic glucose uptake was used as an indicator of epithelial function and was demonstrated by cellular accumulation of tracer (14)C D-glucose and Ussing chamber based short-circuit currents. Assessment of the epithelia by light and immunofluorescent microscopy revealed the harvested enterocytes remain differentiated and establish cell-cell connections to form polarized epithelia with distinct apical and basolateral domains. As with intact tissues, the epithelia exhibit glucose induced short-circuit currents that are increased by exposure to adenosine and adenosine 5'-monophosphate (AMP) and decreased by phloridzin to inhibit the apical glucose transporter SGLT-1. Similarly, accumulation of (14)C D-glucose by the epithelia was inhibited by phloridzin, but not phloretin, and was stimulated by pre-exposure to AMP and adenosine, apparently by a microtubule-based mechanism that is disrupted by nocodazole, with the magnitudes of responses to adenosine, forskolin, and health status exceeding those we have measured using intact tissues. Our findings indicate that epithelia prepared from harvested enterocytes provide an alternative approach for comparative studies of the characteristics of nutrient transport by the upper villus epithelium and the responses to different conditions and stimuli.

U2 - 10.1007/s10616-013-9656-1

DO - 10.1007/s10616-013-9656-1

M3 - Journal article

VL - 67

SP - 39

EP - 49

JO - Cytotechnology

JF - Cytotechnology

SN - 0920-9069

IS - 1

ER -

ID: 46276985