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Generation of insulin‐secreting cells from human embryonic stem cells
Author(s) -
Baharvand Hossein,
Jafary Hanieh,
Massumi Mohammad,
Ashtiani Saeid Kazemi
Publication year - 2006
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/j.1440-169x.2006.00867.x
Subject(s) - embryonic stem cell , stem cell , microbiology and biotechnology , insulin , biology , homeobox protein nanog , endocrinology , induced pluripotent stem cell , genetics , gene
A growth factor‐mediated selection method was used to obtained insulin‐secreting cells from human embryonic stem cells (hESC; Royan H1). Our resultant cells were positive for dithizone, a zinc‐chelating agent known to selectively stain pancreatic β cells and immunoreactive for antibodies against insulin, glucagon, and C‐peptide. Semi‐quantitative reverse transcription–polymerase chain reaction detected expression of proinsulin, insulin and other pancreatic β‐cell‐related genes, such as Nkx6.1, Is11 , Glut2, Pax4 , and prohormone convertase2 ( PC2 ). Moreover, glucagon, somatostatin, K ATP ‐channel genes KIR6.2 and SUR1 , islet amyloid polypeptide ( IAPP ), PC1/3 , and glucokinase ( GCK ) were expressed in the differentiating hESC in a developmental stage‐dependent manner. Also, the addition of glucose to the culture medium triggered insulin release from differentiated cells, but transmission electron microscopy of the differentiated cells did not show typical β‐cell granules, even though secretary granules were detected. The results showed that hESC have the ability to transcribe and process insulin, but further improvements of the current method are required to generate a sufficient source of true β cells for the treatment of diabetes mellitus.