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, Ron 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 journalArticle

4 Citations (Scopus)

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 languageEnglish (US)
JournalXenotransplantation
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Transcriptome
Swine
Insulin
Oxygen
Islets of Langerhans Transplantation
Cell Proliferation
Glucose
Oxygen Consumption
Membranes
RNA Sequence Analysis
Secretory Pathway
Gene Expression Profiling
Type 1 Diabetes Mellitus
Pancreas
In Vitro Techniques
Staining and Labeling
Gene Expression
DNA
Therapeutics

Keywords

  • Islet transplantation
  • Porcine islets
  • RNAseq
  • Type 1 diabetes

ASJC Scopus subject areas

  • Immunology
  • Transplantation

Cite this

In vitro characterization of neonatal, juvenile, and adult porcine islet oxygen demand, β-cell function, and transcriptomes. / Smith, Kate E.; Purvis, William G.; Davis, Melissa A.; Min, Catherine G.; Cooksey, Amanda M.; Weber, Craig S.; Jandova, Jana; Price, Nicholas D.; Molano, Diana S.; Stanton, James Brett; Kelly, Amy C.; Steyn, Leah V.; Lynch, Ron; Limesand, Sean W; Alexander, Michael; Lakey, Jonathan R.T.; Seeberger, Karen; Korbutt, Gregory S.; Mueller, Kate R.; Hering, Bernhard J.; McCarthy, Fiona M; Papas, Klearchos K.

In: Xenotransplantation, 01.01.2018.

Research output: Contribution to journalArticle

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, R, 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. https://doi.org/10.1111/xen.12432
Smith, Kate E. ; Purvis, William G. ; Davis, Melissa A. ; Min, Catherine G. ; Cooksey, Amanda M. ; Weber, Craig S. ; Jandova, Jana ; Price, Nicholas D. ; Molano, Diana S. ; Stanton, James Brett ; Kelly, Amy C. ; Steyn, Leah V. ; Lynch, Ron ; Limesand, Sean W ; Alexander, Michael ; Lakey, Jonathan R.T. ; Seeberger, Karen ; Korbutt, Gregory S. ; Mueller, Kate R. ; Hering, Bernhard J. ; McCarthy, Fiona M ; Papas, Klearchos K. / In vitro characterization of neonatal, juvenile, and adult porcine islet oxygen demand, β-cell function, and transcriptomes. In: Xenotransplantation. 2018.
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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.",
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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, Ron

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

PY - 2018/1/1

Y1 - 2018/1/1

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 - Islet transplantation

KW - Porcine islets

KW - RNAseq

KW - Type 1 diabetes

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