Pancreatic islet composites secrete insulin in response to a glucose challenge

Henry E. Young, Jason Ionnis Limnios, Frank Lochner, George McCommon, Lee Anne Cope, Asa C Black Jr.

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Abstract

This study was designed to test the hypothesis that decellularized pancreatic matrices seeded with adult-derived endogenous stem cells and donor islets provide an optimal environment for islets to secrete insulin in response to a glucose challenge. Adult animals were euthanized following the guidelines of Fort Valley State University-IACUC and Mercer University-IACUC. Adult porcine pancreases were decellularized using a mixture of detergents. Adult rat pancreatic islets were obtained by lipase digestion followed by Ficoll gradient sedimentation. Control cultures consisted of decellularized matrices, clonal populations of naïve adult totipotent and pluripotent stem cell populations, and rat islets, all cultured individually. Experimental groups consisted of islets co-cultured with clonal populations of pluripotent stem cells and totipotent stem cells seeded on decellularized matrices. Control and experimental cultures were challenged with the insulin secretagogue glucose. The control and culture media were removed and stored at -20oC until assayed using a RIA specific for rat insulin. The culture media, containing bovine insulin, were assayed using a RIA specific for rat insulin. No detectable levels of insulin (bovine, rat, human, or porcine) were noted in media only, the stem cell populations or the decellularized matrices, respectively. Native pancreatic islets secreted nanogram quantities of insulin per nanogram of DNA. Pancreatic islets co-cultured with naïve stem cells and matrices demonstrated increased insulin secretion in the range of milligram quantities of insulin per nanogram of DNA, i.e., a 250-fold increase in insulin secretion in response compared to pancreatic islets alone. These studies suggest that native islets in combination with decellularized matrices and adult-derived pluripotent and totipotent stem cells could provide more tissue for pancreatic islet transplants than donor islets alone.
Original languageEnglish
Number of pages12
JournalJournal of Stem Cell Research
Volume1
Issue number1
Publication statusPublished - 17 Apr 2017

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Islets of Langerhans
Insulin
Glucose
Totipotent Stem Cells
Pluripotent Stem Cells
Animal Care Committees
Stem Cells
Population
Culture Media
Swine
Ficoll
Adult Stem Cells
DNA
Lipase
Detergents
Pancreas
Digestion
Tissue Donors
Guidelines

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Young, H. E., Limnios, J. I., Lochner, F., McCommon, G., Cope, L. A., & Black Jr., A. C. (2017). Pancreatic islet composites secrete insulin in response to a glucose challenge. Journal of Stem Cell Research, 1(1).
Young, Henry E. ; Limnios, Jason Ionnis ; Lochner, Frank ; McCommon, George ; Cope, Lee Anne ; Black Jr., Asa C. / Pancreatic islet composites secrete insulin in response to a glucose challenge. In: Journal of Stem Cell Research. 2017 ; Vol. 1, No. 1.
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Young, HE, Limnios, JI, Lochner, F, McCommon, G, Cope, LA & Black Jr., AC 2017, 'Pancreatic islet composites secrete insulin in response to a glucose challenge' Journal of Stem Cell Research, vol. 1, no. 1.

Pancreatic islet composites secrete insulin in response to a glucose challenge. / Young, Henry E.; Limnios, Jason Ionnis; Lochner, Frank; McCommon, George; Cope, Lee Anne; Black Jr., Asa C.

In: Journal of Stem Cell Research, Vol. 1, No. 1, 17.04.2017.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Limnios, Jason Ionnis

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AU - McCommon, George

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N2 - This study was designed to test the hypothesis that decellularized pancreatic matrices seeded with adult-derived endogenous stem cells and donor islets provide an optimal environment for islets to secrete insulin in response to a glucose challenge. Adult animals were euthanized following the guidelines of Fort Valley State University-IACUC and Mercer University-IACUC. Adult porcine pancreases were decellularized using a mixture of detergents. Adult rat pancreatic islets were obtained by lipase digestion followed by Ficoll gradient sedimentation. Control cultures consisted of decellularized matrices, clonal populations of naïve adult totipotent and pluripotent stem cell populations, and rat islets, all cultured individually. Experimental groups consisted of islets co-cultured with clonal populations of pluripotent stem cells and totipotent stem cells seeded on decellularized matrices. Control and experimental cultures were challenged with the insulin secretagogue glucose. The control and culture media were removed and stored at -20oC until assayed using a RIA specific for rat insulin. The culture media, containing bovine insulin, were assayed using a RIA specific for rat insulin. No detectable levels of insulin (bovine, rat, human, or porcine) were noted in media only, the stem cell populations or the decellularized matrices, respectively. Native pancreatic islets secreted nanogram quantities of insulin per nanogram of DNA. Pancreatic islets co-cultured with naïve stem cells and matrices demonstrated increased insulin secretion in the range of milligram quantities of insulin per nanogram of DNA, i.e., a 250-fold increase in insulin secretion in response compared to pancreatic islets alone. These studies suggest that native islets in combination with decellularized matrices and adult-derived pluripotent and totipotent stem cells could provide more tissue for pancreatic islet transplants than donor islets alone.

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JO - Journal of Stem Cell Research

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