In vitro characterization of neonatal, juvenile, and adult porcine islet oxygen demand, β-cell function, and transcriptomes

Kate E. Smith; William G. Purvis; Melissa A. Davis; Catherine G. Min; Amanda M. Cooksey; Craig S. Weber; Jana Jandova; Nicholas D. Price; Diana S. Molano; James Brett Stanton; Amy C. Kelly; Leah V. Steyn; Ronald M. Lynch; Sean W. Limesand; Michael Alexander; Jonathan R.T. Lakey; Karen Seeberger; Gregory S. Korbutt; Kate R. Mueller; Bernhard J. Hering; Fiona M. McCarthy; Klearchos K. Papas

doi:10.1111/xen.12432

In vitro characterization of neonatal, juvenile, and adult porcine islet oxygen demand, β-cell function, and transcriptomes

Kate E. Smith, William G. Purvis, Melissa A. Davis, Catherine G. Min, Amanda M. Cooksey, Craig S. Weber, Jana Jandova, Nicholas D. Price, Diana S. Molano, James Brett Stanton, Amy C. Kelly, Leah V. Steyn, Ronald M. Lynch, Sean W. Limesand, Michael Alexander, Jonathan R.T. Lakey, Karen Seeberger, Gregory S. Korbutt, Kate R. Mueller, Bernhard J. HeringFiona M. McCarthy, Klearchos K. Papas

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

20 Scopus citations

Abstract

Background: There is currently a shortage of human donor pancreata which limits the broad application of islet transplantation as a treatment for type 1 diabetes. Porcine islets have demonstrated potential as an alternative source, but a study evaluating islets from different donor ages under unified protocols has yet to be conducted. Methods: Neonatal porcine islets (NPI; 1-3 days), juvenile porcine islets (JPI; 18-21 days), and adult porcine islets (API; 2+ years) were compared in vitro, including assessments of oxygen consumption rate, membrane integrity determined by FDA/PI staining, β-cell proliferation, dynamic glucose-stimulated insulin secretion, and RNA sequencing. Results: Oxygen consumption rate normalized to DNA was not significantly different between ages. Membrane integrity was age dependent, and API had the highest percentage of intact cells. API also had the highest glucose-stimulated insulin secretion response during a dynamic insulin secretion assay and had 50-fold higher total insulin content compared to NPI and JPI. NPI and JPI had similar glucose responsiveness, β-cell percentage, and β-cell proliferation rate. Transcriptome analysis was consistent with physiological assessments. API transcriptomes were enriched for cellular metabolic and insulin secretory pathways, while NPI exhibited higher expression of genes associated with proliferation. Conclusions: The oxygen demand, membrane integrity, β-cell function and proliferation, and transcriptomes of islets from API, JPI, and NPI provide a comprehensive physiological comparison for future studies. These assessments will inform the optimal application of each age of porcine islet to expand the availability of islet transplantation.

Original language	English (US)
Article number	e12432
Journal	Xenotransplantation
Volume	25
Issue number	6
DOIs	https://doi.org/10.1111/xen.12432
State	Published - Nov 1 2018

Keywords

RNAseq
islet transplantation
porcine islets
type 1 diabetes

ASJC Scopus subject areas

Immunology
Transplantation

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10.1111/xen.12432

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Smith, K. E., Purvis, W. G., Davis, M. A., Min, C. G., Cooksey, A. M., Weber, C. S., Jandova, J., Price, N. D., Molano, D. S., Stanton, J. B., Kelly, A. C., Steyn, L. V., Lynch, R. M., Limesand, S. W., Alexander, M., Lakey, J. R. T., Seeberger, K., Korbutt, G. S., Mueller, K. R., ... Papas, K. K. (2018). In vitro characterization of neonatal, juvenile, and adult porcine islet oxygen demand, β-cell function, and transcriptomes. Xenotransplantation, 25(6), Article e12432. https://doi.org/10.1111/xen.12432

Smith, KE, Purvis, WG, Davis, MA, Min, CG, Cooksey, AM, Weber, CS, Jandova, J, Price, ND, Molano, DS, Stanton, JB, Kelly, AC, Steyn, LV, Lynch, RM , Limesand, SW, Alexander, M, Lakey, JRT, Seeberger, K, Korbutt, GS, Mueller, KR, Hering, BJ, McCarthy, FM & Papas, KK 2018, 'In vitro characterization of neonatal, juvenile, and adult porcine islet oxygen demand, β-cell function, and transcriptomes', Xenotransplantation, vol. 25, no. 6, e12432. https://doi.org/10.1111/xen.12432

@article{1a0bf3e9701f45b4b96b1d04c02f7abd,

title = "In vitro characterization of neonatal, juvenile, and adult porcine islet oxygen demand, β-cell function, and transcriptomes",

abstract = "Background: There is currently a shortage of human donor pancreata which limits the broad application of islet transplantation as a treatment for type 1 diabetes. Porcine islets have demonstrated potential as an alternative source, but a study evaluating islets from different donor ages under unified protocols has yet to be conducted. Methods: Neonatal porcine islets (NPI; 1-3 days), juvenile porcine islets (JPI; 18-21 days), and adult porcine islets (API; 2+ years) were compared in vitro, including assessments of oxygen consumption rate, membrane integrity determined by FDA/PI staining, β-cell proliferation, dynamic glucose-stimulated insulin secretion, and RNA sequencing. Results: Oxygen consumption rate normalized to DNA was not significantly different between ages. Membrane integrity was age dependent, and API had the highest percentage of intact cells. API also had the highest glucose-stimulated insulin secretion response during a dynamic insulin secretion assay and had 50-fold higher total insulin content compared to NPI and JPI. NPI and JPI had similar glucose responsiveness, β-cell percentage, and β-cell proliferation rate. Transcriptome analysis was consistent with physiological assessments. API transcriptomes were enriched for cellular metabolic and insulin secretory pathways, while NPI exhibited higher expression of genes associated with proliferation. Conclusions: The oxygen demand, membrane integrity, β-cell function and proliferation, and transcriptomes of islets from API, JPI, and NPI provide a comprehensive physiological comparison for future studies. These assessments will inform the optimal application of each age of porcine islet to expand the availability of islet transplantation.",

keywords = "RNAseq, islet transplantation, porcine islets, type 1 diabetes",

author = "Smith, {Kate E.} and Purvis, {William G.} and Davis, {Melissa A.} and Min, {Catherine G.} and Cooksey, {Amanda M.} and Weber, {Craig S.} and Jana Jandova and Price, {Nicholas D.} and Molano, {Diana S.} and Stanton, {James Brett} and Kelly, {Amy C.} and Steyn, {Leah V.} and Lynch, {Ronald M.} and Limesand, {Sean W.} and Michael Alexander and Lakey, {Jonathan R.T.} and Karen Seeberger and Korbutt, {Gregory S.} and Mueller, {Kate R.} and Hering, {Bernhard J.} and McCarthy, {Fiona M.} and Papas, {Klearchos K.}",

note = "Funding Information: This research was supported by JDRF grant #2-SRA-2014-289-Q-R and by the National Institutes of Health Interdisciplinary Training Grant in Cardiovascular Sciences (HL007249). JPI supply was supported by JDRF grant #17-2013-288. Funding Information: Funding information This research was supported by JDRF grant #2-SRA- 2014-289- Q-R and by the National Institutes of Health Interdisciplinary Training Grant in Cardiovascular Sciences (HL007249). JPI supply was supported by JDRF grant #17-2013-288. We would like to acknowledge the members of the Papas Laboratory, including Anna Maria Burachek, Alexandra Hoeger, Erik Rohner, and Ivan Georgiev. We would also like to thank the University of Arizona Genetics Core, especially Jonathan Galina-Mehlman, and the ARCS Foundation Phoenix Chapter for their support. Publisher Copyright: {\textcopyright} 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.",

year = "2018",

month = nov,

day = "1",

doi = "10.1111/xen.12432",

language = "English (US)",

volume = "25",

journal = "Xenotransplantation",

issn = "0908-665X",

publisher = "Wiley-Blackwell",

number = "6",

}

TY - JOUR

T1 - In vitro characterization of neonatal, juvenile, and adult porcine islet oxygen demand, β-cell function, and transcriptomes

AU - Smith, Kate E.

AU - Purvis, William G.

AU - Davis, Melissa A.

AU - Min, Catherine G.

AU - Cooksey, Amanda M.

AU - Weber, Craig S.

AU - Jandova, Jana

AU - Price, Nicholas D.

AU - Molano, Diana S.

AU - Stanton, James Brett

AU - Kelly, Amy C.

AU - Steyn, Leah V.

AU - Lynch, Ronald M.

AU - Limesand, Sean W.

AU - Alexander, Michael

AU - Lakey, Jonathan R.T.

AU - Seeberger, Karen

AU - Korbutt, Gregory S.

AU - Mueller, Kate R.

AU - Hering, Bernhard J.

AU - McCarthy, Fiona M.

AU - Papas, Klearchos K.

N1 - Funding Information: This research was supported by JDRF grant #2-SRA-2014-289-Q-R and by the National Institutes of Health Interdisciplinary Training Grant in Cardiovascular Sciences (HL007249). JPI supply was supported by JDRF grant #17-2013-288. Funding Information: Funding information This research was supported by JDRF grant #2-SRA- 2014-289- Q-R and by the National Institutes of Health Interdisciplinary Training Grant in Cardiovascular Sciences (HL007249). JPI supply was supported by JDRF grant #17-2013-288. We would like to acknowledge the members of the Papas Laboratory, including Anna Maria Burachek, Alexandra Hoeger, Erik Rohner, and Ivan Georgiev. We would also like to thank the University of Arizona Genetics Core, especially Jonathan Galina-Mehlman, and the ARCS Foundation Phoenix Chapter for their support. Publisher Copyright: © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

PY - 2018/11/1

Y1 - 2018/11/1

N2 - Background: There is currently a shortage of human donor pancreata which limits the broad application of islet transplantation as a treatment for type 1 diabetes. Porcine islets have demonstrated potential as an alternative source, but a study evaluating islets from different donor ages under unified protocols has yet to be conducted. Methods: Neonatal porcine islets (NPI; 1-3 days), juvenile porcine islets (JPI; 18-21 days), and adult porcine islets (API; 2+ years) were compared in vitro, including assessments of oxygen consumption rate, membrane integrity determined by FDA/PI staining, β-cell proliferation, dynamic glucose-stimulated insulin secretion, and RNA sequencing. Results: Oxygen consumption rate normalized to DNA was not significantly different between ages. Membrane integrity was age dependent, and API had the highest percentage of intact cells. API also had the highest glucose-stimulated insulin secretion response during a dynamic insulin secretion assay and had 50-fold higher total insulin content compared to NPI and JPI. NPI and JPI had similar glucose responsiveness, β-cell percentage, and β-cell proliferation rate. Transcriptome analysis was consistent with physiological assessments. API transcriptomes were enriched for cellular metabolic and insulin secretory pathways, while NPI exhibited higher expression of genes associated with proliferation. Conclusions: The oxygen demand, membrane integrity, β-cell function and proliferation, and transcriptomes of islets from API, JPI, and NPI provide a comprehensive physiological comparison for future studies. These assessments will inform the optimal application of each age of porcine islet to expand the availability of islet transplantation.

AB - Background: There is currently a shortage of human donor pancreata which limits the broad application of islet transplantation as a treatment for type 1 diabetes. Porcine islets have demonstrated potential as an alternative source, but a study evaluating islets from different donor ages under unified protocols has yet to be conducted. Methods: Neonatal porcine islets (NPI; 1-3 days), juvenile porcine islets (JPI; 18-21 days), and adult porcine islets (API; 2+ years) were compared in vitro, including assessments of oxygen consumption rate, membrane integrity determined by FDA/PI staining, β-cell proliferation, dynamic glucose-stimulated insulin secretion, and RNA sequencing. Results: Oxygen consumption rate normalized to DNA was not significantly different between ages. Membrane integrity was age dependent, and API had the highest percentage of intact cells. API also had the highest glucose-stimulated insulin secretion response during a dynamic insulin secretion assay and had 50-fold higher total insulin content compared to NPI and JPI. NPI and JPI had similar glucose responsiveness, β-cell percentage, and β-cell proliferation rate. Transcriptome analysis was consistent with physiological assessments. API transcriptomes were enriched for cellular metabolic and insulin secretory pathways, while NPI exhibited higher expression of genes associated with proliferation. Conclusions: The oxygen demand, membrane integrity, β-cell function and proliferation, and transcriptomes of islets from API, JPI, and NPI provide a comprehensive physiological comparison for future studies. These assessments will inform the optimal application of each age of porcine islet to expand the availability of islet transplantation.

KW - RNAseq

KW - islet transplantation

KW - porcine islets

KW - type 1 diabetes

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UR - http://www.scopus.com/inward/citedby.url?scp=85051051129&partnerID=8YFLogxK

U2 - 10.1111/xen.12432

DO - 10.1111/xen.12432

M3 - Article

C2 - 30052287

AN - SCOPUS:85051051129

SN - 0908-665X

VL - 25

JO - Xenotransplantation

JF - Xenotransplantation

IS - 6

M1 - e12432

ER -

In vitro characterization of neonatal, juvenile, and adult porcine islet oxygen demand, β-cell function, and transcriptomes

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