Co-axial Printing of Convoluted Proximal Tubule for Kidney Disease Modeling

Anne Metje van Genderen; Marta G Valverde; Pamela E Capendale; Valerie Kersten; Elena Sendino Garví; Carl C L Schuurmans; Marina Ruelas; Joana T Soeiro; Guosheng Tang; Manoe J Janssen; Jitske Jansen; Silvia M Mihăilă; Tina Vermonden; Y Shrike Zhang; Rosalinde Masereeuw

doi:10.1088/1758-5090/ac7895

Co-axial Printing of Convoluted Proximal Tubule for Kidney Disease Modeling

Anne Metje van Genderen, Marta G Valverde, Pamela E Capendale, Valerie Kersten, Elena Sendino Garví, Carl C L Schuurmans, Marina Ruelas, Joana T Soeiro, Guosheng Tang, Manoe J Janssen, Jitske Jansen, Silvia M Mihăilă, Tina Vermonden, Y Shrike Zhang, Rosalinde Masereeuw

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Despite the increasing incidence of kidney-related diseases, we are still far from understanding the underlying mechanisms of these diseases and their progression. This lack of understanding is partly because of a poor replication of the diseasesin vitro,limited to planar culture. Advancing towards three-dimensional models, hereby we propose coaxial printing to obtain microfibers containing a helical hollow microchannel. These recapitulate the architecture of the proximal tubule (PT), an important nephron segment often affected in kidney disorders. A stable gelatin/alginate-based ink was formulated to allow printability while maintaining structural properties. Fine-tuning of the composition, printing temperature and extrusion rate allowed for optimal ink viscosity that led to coiling of the microfiber's inner channel. The printed microfibers exhibited prolonged structural stability (42 days) and cytocompatibility in culture. Healthy conditionally immortalized PT epithelial cells and a knockout cell model for cystinosis (CTNS-/-) were seeded to mimic two genotypes of PT. Upon culturing for 14 days, engineered PT showed homogenous cytoskeleton organization as indicated by staining for filamentous actin, barrier-formation and polarization with apical markerα-tubulin and basolateral marker Na+/K+-ATPase. Cell viability was slightly decreased upon prolonged culturing for 14 days, which was more pronounced inCTNS-/-microfibers. Finally,CTNS-/-cells showed reduced apical transport activity in the microfibers compared to healthy PT epithelial cells when looking at breast cancer resistance protein and multidrug resistance-associated protein 4. Engineered PT incorporated in a custom-designed microfluidic chip allowed to assess leak-tightness of the epithelium, which appeared less tight inCTNS-/-PT compared to healthy PT, in agreement with itsin vivophenotype. While we are still on the verge of patient-oriented medicine, this system holds great promise for further research in establishing advancedin vitrodisease models.

Original language	English
Article number	044102
Pages (from-to)	1-16
Journal	Biofabrication
Volume	14
Issue number	4
Early online date	14 Jun 2022
DOIs	https://doi.org/10.1088/1758-5090/ac7895
Publication status	Published - Oct 2022

Bibliographical note

Funding Information:
This work was supported by funding from The Dutch Kidney Foundation (17PHD16, A M G and 18KVP01, A M G, C S, Y S Z), the Hofvijverkring Visiting Scientist Program (Y S Z, A M G, C S), the Materials Driven Regeneration Young Talent Grant (A M G, C S), H2020 WIDESPREAD-05-2018-TWINNING Remodel (S M M; R M), the IMAGEN project which is co-funded by the PPP Allowance made available by Health∼Holland, Top Sector Life Sciencezs & Health, to stimulate public–private partnerships (IMplementation of Advancements in GENetic Kidney Disease, LSHM20009; E S G, R M), Utrecht Institute for Pharmaceutical Sciences (M G V) and Brigham Research Institute (Y S Z).

Publisher Copyright:
© 2022 The Author(s). Published by IOP Publishing Ltd.

Keywords

coaxial 3D printing
biomaterials
proximal tubule
cystinosis
in vitro modeling
kidney

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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van_Genderen_2022_Biofabrication_14_044102Final published version, 3.31 MBLicence: CC BY

Cite this

Genderen, A. M. V., G Valverde, M., Capendale, P. E., Kersten, V., Sendino Garví, E., Schuurmans, C. C. L., Ruelas, M., Soeiro, J. T., Tang, G., Janssen, M. J., Jansen, J., Mihăilă, S. M., Vermonden, T., Zhang, Y. S., & Masereeuw, R. (2022). Co-axial Printing of Convoluted Proximal Tubule for Kidney Disease Modeling. Biofabrication, 14(4), 1-16. Article 044102. https://doi.org/10.1088/1758-5090/ac7895

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abstract = "Despite the increasing incidence of kidney-related diseases, we are still far from understanding the underlying mechanisms of these diseases and their progression. This lack of understanding is partly because of a poor replication of the diseasesin vitro,limited to planar culture. Advancing towards three-dimensional models, hereby we propose coaxial printing to obtain microfibers containing a helical hollow microchannel. These recapitulate the architecture of the proximal tubule (PT), an important nephron segment often affected in kidney disorders. A stable gelatin/alginate-based ink was formulated to allow printability while maintaining structural properties. Fine-tuning of the composition, printing temperature and extrusion rate allowed for optimal ink viscosity that led to coiling of the microfiber's inner channel. The printed microfibers exhibited prolonged structural stability (42 days) and cytocompatibility in culture. Healthy conditionally immortalized PT epithelial cells and a knockout cell model for cystinosis (CTNS-/-) were seeded to mimic two genotypes of PT. Upon culturing for 14 days, engineered PT showed homogenous cytoskeleton organization as indicated by staining for filamentous actin, barrier-formation and polarization with apical markerα-tubulin and basolateral marker Na+/K+-ATPase. Cell viability was slightly decreased upon prolonged culturing for 14 days, which was more pronounced inCTNS-/-microfibers. Finally,CTNS-/-cells showed reduced apical transport activity in the microfibers compared to healthy PT epithelial cells when looking at breast cancer resistance protein and multidrug resistance-associated protein 4. Engineered PT incorporated in a custom-designed microfluidic chip allowed to assess leak-tightness of the epithelium, which appeared less tight inCTNS-/-PT compared to healthy PT, in agreement with itsin vivophenotype. While we are still on the verge of patient-oriented medicine, this system holds great promise for further research in establishing advancedin vitrodisease models.",

keywords = "coaxial 3D printing, biomaterials, proximal tubule, cystinosis, in vitro modeling, kidney",

author = "Genderen, {Anne Metje van} and {G Valverde}, Marta and Capendale, {Pamela E} and Valerie Kersten and {Sendino Garv{\'i}}, Elena and Schuurmans, {Carl C L} and Marina Ruelas and Soeiro, {Joana T} and Guosheng Tang and Janssen, {Manoe J} and Jitske Jansen and Mih{\u a}il{\u a}, {Silvia M} and Tina Vermonden and Zhang, {Y Shrike} and Rosalinde Masereeuw",

note = "Funding Information: This work was supported by funding from The Dutch Kidney Foundation (17PHD16, A M G and 18KVP01, A M G, C S, Y S Z), the Hofvijverkring Visiting Scientist Program (Y S Z, A M G, C S), the Materials Driven Regeneration Young Talent Grant (A M G, C S), H2020 WIDESPREAD-05-2018-TWINNING Remodel (S M M; R M), the IMAGEN project which is co-funded by the PPP Allowance made available by Health∼Holland, Top Sector Life Sciencezs & Health, to stimulate public–private partnerships (IMplementation of Advancements in GENetic Kidney Disease, LSHM20009; E S G, R M), Utrecht Institute for Pharmaceutical Sciences (M G V) and Brigham Research Institute (Y S Z). Publisher Copyright: {\textcopyright} 2022 The Author(s). Published by IOP Publishing Ltd.",

year = "2022",

month = oct,

doi = "10.1088/1758-5090/ac7895",

language = "English",

volume = "14",

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issn = "1758-5082",

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Genderen, AMV, G Valverde, M, Capendale, PE, Kersten, V, Sendino Garví, E, Schuurmans, CCL, Ruelas, M, Soeiro, JT, Tang, G, Janssen, MJ, Jansen, J, Mihăilă, SM , Vermonden, T, Zhang, YS & Masereeuw, R 2022, 'Co-axial Printing of Convoluted Proximal Tubule for Kidney Disease Modeling', Biofabrication, vol. 14, no. 4, 044102, pp. 1-16. https://doi.org/10.1088/1758-5090/ac7895

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T1 - Co-axial Printing of Convoluted Proximal Tubule for Kidney Disease Modeling

AU - Genderen, Anne Metje van

AU - G Valverde, Marta

AU - Capendale, Pamela E

AU - Kersten, Valerie

AU - Sendino Garví, Elena

AU - Schuurmans, Carl C L

AU - Ruelas, Marina

AU - Soeiro, Joana T

AU - Tang, Guosheng

AU - Janssen, Manoe J

AU - Jansen, Jitske

AU - Mihăilă, Silvia M

AU - Vermonden, Tina

AU - Zhang, Y Shrike

AU - Masereeuw, Rosalinde

N1 - Funding Information: This work was supported by funding from The Dutch Kidney Foundation (17PHD16, A M G and 18KVP01, A M G, C S, Y S Z), the Hofvijverkring Visiting Scientist Program (Y S Z, A M G, C S), the Materials Driven Regeneration Young Talent Grant (A M G, C S), H2020 WIDESPREAD-05-2018-TWINNING Remodel (S M M; R M), the IMAGEN project which is co-funded by the PPP Allowance made available by Health∼Holland, Top Sector Life Sciencezs & Health, to stimulate public–private partnerships (IMplementation of Advancements in GENetic Kidney Disease, LSHM20009; E S G, R M), Utrecht Institute for Pharmaceutical Sciences (M G V) and Brigham Research Institute (Y S Z). Publisher Copyright: © 2022 The Author(s). Published by IOP Publishing Ltd.

PY - 2022/10

Y1 - 2022/10

N2 - Despite the increasing incidence of kidney-related diseases, we are still far from understanding the underlying mechanisms of these diseases and their progression. This lack of understanding is partly because of a poor replication of the diseasesin vitro,limited to planar culture. Advancing towards three-dimensional models, hereby we propose coaxial printing to obtain microfibers containing a helical hollow microchannel. These recapitulate the architecture of the proximal tubule (PT), an important nephron segment often affected in kidney disorders. A stable gelatin/alginate-based ink was formulated to allow printability while maintaining structural properties. Fine-tuning of the composition, printing temperature and extrusion rate allowed for optimal ink viscosity that led to coiling of the microfiber's inner channel. The printed microfibers exhibited prolonged structural stability (42 days) and cytocompatibility in culture. Healthy conditionally immortalized PT epithelial cells and a knockout cell model for cystinosis (CTNS-/-) were seeded to mimic two genotypes of PT. Upon culturing for 14 days, engineered PT showed homogenous cytoskeleton organization as indicated by staining for filamentous actin, barrier-formation and polarization with apical markerα-tubulin and basolateral marker Na+/K+-ATPase. Cell viability was slightly decreased upon prolonged culturing for 14 days, which was more pronounced inCTNS-/-microfibers. Finally,CTNS-/-cells showed reduced apical transport activity in the microfibers compared to healthy PT epithelial cells when looking at breast cancer resistance protein and multidrug resistance-associated protein 4. Engineered PT incorporated in a custom-designed microfluidic chip allowed to assess leak-tightness of the epithelium, which appeared less tight inCTNS-/-PT compared to healthy PT, in agreement with itsin vivophenotype. While we are still on the verge of patient-oriented medicine, this system holds great promise for further research in establishing advancedin vitrodisease models.

AB - Despite the increasing incidence of kidney-related diseases, we are still far from understanding the underlying mechanisms of these diseases and their progression. This lack of understanding is partly because of a poor replication of the diseasesin vitro,limited to planar culture. Advancing towards three-dimensional models, hereby we propose coaxial printing to obtain microfibers containing a helical hollow microchannel. These recapitulate the architecture of the proximal tubule (PT), an important nephron segment often affected in kidney disorders. A stable gelatin/alginate-based ink was formulated to allow printability while maintaining structural properties. Fine-tuning of the composition, printing temperature and extrusion rate allowed for optimal ink viscosity that led to coiling of the microfiber's inner channel. The printed microfibers exhibited prolonged structural stability (42 days) and cytocompatibility in culture. Healthy conditionally immortalized PT epithelial cells and a knockout cell model for cystinosis (CTNS-/-) were seeded to mimic two genotypes of PT. Upon culturing for 14 days, engineered PT showed homogenous cytoskeleton organization as indicated by staining for filamentous actin, barrier-formation and polarization with apical markerα-tubulin and basolateral marker Na+/K+-ATPase. Cell viability was slightly decreased upon prolonged culturing for 14 days, which was more pronounced inCTNS-/-microfibers. Finally,CTNS-/-cells showed reduced apical transport activity in the microfibers compared to healthy PT epithelial cells when looking at breast cancer resistance protein and multidrug resistance-associated protein 4. Engineered PT incorporated in a custom-designed microfluidic chip allowed to assess leak-tightness of the epithelium, which appeared less tight inCTNS-/-PT compared to healthy PT, in agreement with itsin vivophenotype. While we are still on the verge of patient-oriented medicine, this system holds great promise for further research in establishing advancedin vitrodisease models.

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KW - biomaterials

KW - proximal tubule

KW - cystinosis

KW - in vitro modeling

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ER -

Co-axial Printing of Convoluted Proximal Tubule for Kidney Disease Modeling

Abstract

Bibliographical note

Keywords

UN SDGs

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