Embryonic stem cells differentiate into insulin-producing cells without selection of nestin-expressing cells

Measurement of T1, T2, and magnetization transfer properties during embryonic development at 7 Tesla using the chicken model

“…Embryonic stem cells can produce derivatives of all three primary germ cell layers: ectoderm, mesoderm, and endoderm. Numerous differentiation protocols have already been established permitting the generation of almost any somatic cell type such as cardiomyocytes for cardiac repair (5), neural precursor cells with the potential to repair or limit the damage associated with infarct or neurodegenerative diseases (6), or ␤-islet progenitor cells producing insulin for the treatment of diabetes (7,8).…”

Generation of pancreatic insulin-producing cells from embryonic stem cells — ‘Proof of principle’, but questions still unanswered

“…Instead, using the same differentiation protocol, it was found that insulin immunoreactivity occurred as a consequence of insulin uptake from the medium [2], neuronal cells were formed [2][3][4], or insulin was released as an artefact from differentiated ES cells [2,3]. Functional pancreatic cells, however, were successfully generated using lineage selection strategies based on pancreas-specific promoters [5,6], by modified protocols in combination with transgene expression [7][8][9], or by addition of a phosphoinositol-3 kinase inhibitor [10]. The differentiated cells showed properties of (neonatal) beta cells, such as insulin transcripts and C-peptide/insulin co-expression, insulin-secretory granules, ion channel activity of embryonal beta cells, and normalisation of blood glucose level after transplantation into diabetic mice [5][6][7][8][9][10].…”

“…Functional pancreatic cells, however, were successfully generated using lineage selection strategies based on pancreas-specific promoters [5,6], by modified protocols in combination with transgene expression [7][8][9], or by addition of a phosphoinositol-3 kinase inhibitor [10]. The differentiated cells showed properties of (neonatal) beta cells, such as insulin transcripts and C-peptide/insulin co-expression, insulin-secretory granules, ion channel activity of embryonal beta cells, and normalisation of blood glucose level after transplantation into diabetic mice [5][6][7][8][9][10]. Most of the differentiation protocols required a long cultivation period, including 4-5 days of embryoid body (EB) formation, followed by 3-4 weeks of differentiation, and some protocols required genetic manipulation.…”

Differentiation of Stem Cells into Insulin-Producing Cells: Current Status and Challenges

“…Boyd et al (2008) compared three different protocols to differentiate ESCs into IPCs using the embryoid body formation strategy. This study indicated that the IPCs generated using Blyszczuk protocol (Blyszczuk et al 2004) exhibited superior C-peptide expression and longer normoglycemic rescue in diabetic mice when compared with the Hori's and Lumelsky's protocols (Hori et al 2002;Lumelsky et al 2001). In protocol described by Blyszczuk the ESCs were cultured in ''handing drops'' and in suspension for 5 days to form embryoid bodies.…”

“…Then, embryoid bodies were plated in Iscove medium supplemented with 20 % fetal calf serum (FCS), L-glutamine, non-essential amino acids and a-monothioglycerol and cultured for 9 days for differentiation into cells representing derivatives of all three primary germ layers. In the next step, cells were re-plated onto poly-L-ornithine/laminin-coated culture dishes into a pancreatic differentiation medium (N2 medium supplemented with progesterone, putrescine, laminin (LAM), insulin, sodium selenite, nicotinamide, transferrin, fibronectin (FN), B27 media supplement, 15 % FCS) and cultured for 19 days (Blyszczuk et al 2004). The Lumelsky's protocol involves serum-free ITSFn medium and basic fibroblast growth factor (bFGF) treatments (Lumelsky et al 2001).…”