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Improvement of isolated caprine islet survival and functionality in vitro by enhancing of PDX 1 gene expression
Author(s) -
Hani Homayoun,
Allaudin Zeenathul Nazariah,
MohdLila MohdAzmi,
Sarsaifi Kazhal,
Rasouli Mina,
Tam Yew Joon,
TengkuIbrahim TengkuAzmi,
Othman Abas Mazni
Publication year - 2017
Publication title -
xenotransplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.052
H-Index - 61
eISSN - 1399-3089
pISSN - 0908-665X
DOI - 10.1111/xen.12302
Subject(s) - pdx1 , islet , ibmx , insulin , endocrinology , medicine , pancreas , biology , gene expression , chemistry , transplantation , andrology , gene , biochemistry , stimulation , forskolin
Background Dead islets replaced with viable islets are a promising offer to restore normal insulin production to a person with diabetes. The main reason for establishing a new islet source for transplantation is the insufficiency of human donor pancreas while using xenogeneic islets perhaps assists this problem. The expression of PDX 1 is essential for the pancreas expansion. In mature β‐cells, PDX 1 has several critical roles such as glucose sensing, insulin synthesis, and insulin secretion. In this study, we aimed to evaluate the expression of pancreatic duodenal homeobox‐1 ( PDX 1) in treated caprine islets in culture and to assess the protective effects of antioxidant factors on the PDX 1 gene in cultured caprine islets. Materials and methods Purified islets were treated with serum‐free, serum, IBMX , tocopherol, or IBMX and tocopherol media. Quantitative polymerase chain reaction and Western blotting were carried out to compare the expression levels of PDX 1 in treated purified islets cultured with different media. Results Islets treated with IBMX /tocopherol exhibited the highest fold change in the relative expression of PDX 1 on day 5 post‐treatment (relative expression: 6.80±2.08), whereas serum‐treated islets showed the lowest fold changes in PDX 1 expression on day 5 post‐treatment (0.67±0.36), as compared with the expression on day 1 post‐treatment. Insulin production and viability tests of purified islets showed superiority of islet at supplemented serum‐free media with IBMX /tocopherol compared to other cultures (53.875%±1.59%). Conclusions Our results indicated that supplemented serum‐free medium with tocopherol and IBMX enhances viability and PDX 1 gene expression compared to serum‐added and serum‐free media.