Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo

“endocrine cells after implantation into mice (226).”

“, 2013), thereby inspiring the development of protocols for the generation of pancreatic progenitors (Kroon et al., 2008) and subsequently b-like cells from human pluripotent stem cells (Pagliuca et al.”

“Prevailing strategies typically differentiate hPSC-derived endoderm into “primitive gut tube”, “posterior foregut,” and then “pancreatic progenitors” (Kroon et al., 2008; Pagliuca et al., 2014; Rezania et al., 2014) (Figure 2bi).”

“Upon transplantation into immunocompromised mice, the graft (composed of endocrine and ductal cells) restored normoglycemia within 2 [66] or 6 weeks [67], a tremendous improvement compared with the 2- to 3-month period required after transplantation of ESCderived pancreatic progenitors [61].”

“For example, transplantation of muscle satellite cells has been shown to lead to functional recovery of dystrophic muscle (Zammit et al., 2006), and pancreatic precursors made from embryonic stem cells can cure diabetic animals (Kroon et al., 2008).”

“While the viability, and hence, developmental capacity of hPSC‐derived pancreatic progenitor, immune‐protected during the longer maturation period in vivo has been demonstrated in rodents [9,30,31], it is yet to be determined in humans.”

“…be efficiently differentiated to pancreatic endoderm (D’Amour et al., 2005, 2006) that forms beta cells following implantation in immune-deficient mice (Kroon et al., 2008; Rezania et al., 2012) or following further in vitro steps (Pagliuca et al., 2014; Rezania et al., 2014; Russ et al., 2015).”

“In vitro differentiation followed the method of Kroon et al. (2008).”

“This retrievable and vascularizing capsule will contain pancreatic progenitor cells that are expected to differentiate into functioning islet cells(29-32, 110).”

“An alternative approach, not yet feasible, is to create new β-cells (2), either by stepwise differentiation of undifferentiated stem or stem-like cells (3), or by transdifferentiation (4), the heritable change of cell identity to an insulin-producing (β-like) cell.”

“When AR/AL level increased from C1 to C2, the circuit’s quadrastability becomes well pronounced, illustrated as four similar-sized colored spheres on the same gray plane, which represents the low-low, GFP, mCherry, and high-high state, respectively (Figure 3—figure supplement 1).”

“Recently, it has been shown that hES cell-derived pancreatic endoderm cells that were transplanted into immunodeficient mice underwent further differentiation and maturation into glucose-responsive INS-secreting cells (Kroon et al., 2008; Rezania et al., 2012, 2013), suggesting that the pancreatic precursors obtained…”

“Similarly, a previous study demonstrated that transplantation of hESCderived pancreatic endoderm in mice could generate insulin-producing cells in vivo [41].”

“Diabetes 59:2265–2270, 2010 Investigators are working to develop functional human -cells from adult or embryonic stem cells (ESCs) or from induced pluripotent stem (iPS) cells (1–4), and human ESC-derived -cells can restore euglycemia in streptozotocin (STZ) diabetic immunodeficient mice (5).”

“Some preclinical studies and clinical trials confirmed the potential of stem cell therapy in treating several diseases, such as diabetes mellitus [26,27], heart failure [28,29], disorders of the nervous system [30–32], and so on.”

“Other studies made use of RA combined with FGF ligands (FGF4, FGF7, FGF10) or of Noggin combined with FGF2 and EGF to induce pancreatic cells albeit at a lower efficiency than for combined Noggin and RA (Table 3) (13,14,16,17,19,23).”

“Based largely on evidence from preclinical studies in immunodeficient mice (Kroon et al., 2008; Kelly et al., 2011; Rezania et al., 2012, 2013; Bruin et al., 2013; Motté et al., 2014), the safety of hESC-derived pancreatic progenitor cells is now being evaluated clinically in a small cohort of…”

“An important recent report shows that when transplanted into mice, human ES-derived cells with a phenotype corresponding to pancreatic endoderm matured to yield cells capable of maintaining near-normal regulation of blood sugar [Kroon et al., 2008].”

“Many researchers have reported differentiation procedures for pancreatic cell lineages from ES cells (Jiang et al., 2007; Kroon et al., 2008).”

“Results showed that threemonths post-transplantation the implant contained glucose-responsive insulinsecreting cells that were functionally very similar to transplanted adult human islets (Kroon et al., 2008).”

“Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan, the ¶Department of Medicine, Division of Diabetes, Endocrinology, and Metabolism, Vanderbilt University Medical Center, Nashville, Tennessee 37232-6303, and the Department of Molecular Physiology & Biophysics, Vanderbilt University Medical School, Nashville, Tenne…”

“Regulated insulin secretion is a critical feature of pancreatic β -cells that has proved difficult to replicate in ES cell-derived insulin-producing cells 2 .”

“In addition, iPS cells should have a safeguard system because iPS cells with teratoma-forming potential can persist even after differentiation, leading to unexpected and undesired events [16].”

“Transplantation of immature cells under the kidney capsule allows for completion of the β cell differentiation pathway in vivo from these stem cell-like progenitors (Kroon et al., 2008).”

“sources for beta cell replacement therapies [1,2,3].”

“Comparison of the mouse and human pluripotent networks revealed, quite surprisingly, that species-specific transposable elements have considerably altered the transcriptional pluripotent circuitry, so that in each species the same core factors bind a distinct set of TF-binding sites and play distinct roles in pluripot…”

“These conclusions are not unequivocal as hypoglycemia might also have arisen from the absent first-phase insulin release mentioned earlier and/or reduced counter-regulatory responses [48].”

“Implanted pancreatic precursor cells derived from hESCs without a scaffold have been shown to reverse hyperglycemia in nude mice, but take several weeks to have an effect.(28,29) Combining these cells with a scaffold may be able to shorten the maturation time needed to achieve functional implants.”

“A later study employed a similar 4-step protocol to obtain pancreatic endoderm cells (Kroon et al., 2008).”

“Current methods involve undefined animal products such as feeders, FBS and Matrigel and only allow for the generation of heterogeneous populations of cells, thus increasing the risk of teratoma formation after transplantation [7, 8].”

“These results support the notion that the generation of mature and functional differentiated cells from hPSC cultures is challenging for many cell types, such as hepatocytes, pancreatic endocrine cells, and cardiomyocytes.((38-40)) In this context, the development of adequate methodology necessary to assess the differ…”

“patterning to endodermal gut tube [5,6], and retinoic acid”

“Through the application of knowledge learned from embryonic development, conditions have been developed to differentiate ESCs/iPSCs in a stepwise manner into desired cell populations (63), including dopamine neurons (64), retinal pigment epithelium (65), floor plate cells (66), hematopoietic cells (67), endothelial ce…”

“Under the appropriate influences, ESCs are able to differentiate into insulin-producing cells [3–11].”

“The combined effect of cell death and de-differentiation may explain the inconsistent results with SCIPC grafts and the long duration needed to achieve graft function reported previously (Kroon et al., 2008; Matveyenko et al., 2010).”

“Human ESC (hESC) has been demonstrated to differentiate into functional β-cells in vivo [76].”

“…organlike structures from hPSCs, including intestine, stomach, lung, kidney, brain and retina among others (D’Amour et al., 2006; Dye et al., 2015; Kroon et al., 2008; Lancaster et al., 2013; McCracken et al., 2014; Nakano et al., 2012; Si-Tayeb et al., 2010; Takasato et al., 2015; Wells and…”

“Embryonic and induced pluripotent stem cells It has already been shown that human embryonic stem cells (ESCs) can be directed to become fully mature beta cells.”

“Virtually all differentiation protocols from either ES or EP cells utilize similar inductive signals at this stage including FGF, inhibition of sonic hedgehog, and RA signaling (Cheng et al., 2012; Kroon et al., 2008; Nostro et al., 2011; Rezania et al., 2013).”

“Heretofore, robust glucose-induced insulin secretion, the main function of b cell, has only been possible in hESC-derived cells after in vitro transplantation for several months (Kroon et al., 2008; D’Amour et al., 2006).”

“…optimism about the possibility that methods currently under development will produce cells resembling pancreatic or islet progenitors from pluripotent cells like human ESCs or iPSCs that can be used to replace b cells destroyed in T1DM (D’Amour et al., 2006; Kroon et al., 2008; Zhang et al., 2009).”

“These data indicate that in vivo , the NS-and Ft-derived PPs give rise to non-pancreatic tissues from multiple germ layers, characteristic of teratoma (Kroon et al., 2008).”

“ES cells differentiated into PP (Figure 3) and transplanted into mice protected against streptozotocin-induced hyperglycemia and showed serum C-peptide that was comparable to 3000 IEQ human islets [127].”

“Different protocols were used to generate cells of the definitive endoderm germ layer and the primitive gut tube: classical induction of DE and primitive gut tubewas conducted as described earlier [9] using a sequential approach with 25 ng/ml wnt3a and 100 ng/ml activin A for one day, followed by a 2 day treatment wit…”

“Modified pancreatic differentiation by Kroon et al. (2008) reported the production of insulin-producing cells from hESC which were also in vivo responsive to glucose.”

“More recently, the Novocell group has applied their protocol to generate endocrine precursors in vitro, and then grafted these progenitors into diabetic mice to promote their full maturation (Kroon et al. 2008), generating cells that secreted insulin at levels comparable to mature b-cells.”

“An alternative approach seeks not to generate mature cells, but uses early definitive endoderm or pancreatic progenitor cells and relies strongly on self-directed differentiation and maturation in order to obtain a functional stem cell-derived beta cell mass in vivo [70,87,88].”

“5, the time point of gene array analysis, we tested the effects of the combined mesenchymal factors on primitive gut tube cells generated via the standard protocol (factors were added after day 6 of the standard protocol) (10).”

“Several efforts have been reported with varying degrees of success (7, 10, 11, 13, 24).”

“studies, which reduces the usage of animal experiments and fills the gap between animal studies and clinical trials to some extent [81].”

“Furthermore, we added Noggin (an inhibitor of bone morphogenetic protein signaling), retinoic acid, and fi-broblast growth factor 7 (FGF7 or KGF) [34–37] sequentially to the cell culture media to differentiate DE cells into pancreatic progenitors.”

“[114] Kroon E, Martinson LA, Kadoya K, Bang AG, Kelly OG, Eliazer S, et al.”

“Our DE-inducing conditions gave rise to pancreatic progenitor markers SOX9 and PDX1 comparably with Kroon et al. (2008) in two different hPSC lines (Figure S4B), demonstrating equivalence in terms of their differentiation potential.”

“…these initial studies, many laboratories have published protocols that promote the production of insulin-expressing beta-like cells from stem cells (Bruin et al., 2013; D’Amour et al., 2006; Kroon et al., 2008; McGrath et al., 2015; Pagliuca et al., 2014; Rezania et al., 2013; Toyoda et al., 2015).”

“…a specific lineage (Tabar and Studer, 2014), with dozens of these applied to in vitro disease modeling, and many fewer to in vivo regeneration (Darabi et al., 2012; Hargus et al., 2010; Kriks et al., 2011; Kroon et al., 2008; Laflamme et al., 2007; Sebastiano et al., 2014; Wang et al., 2012).”

“Although significant progress has been made in the derivation of β-like cells from ES (D’Amour et al., 2006; Jiang et al., 2007; Kroon et al., 2008; Micallef et al., 2012; Nostro et al., 2011; Zhang et al., 2009) and iPS (Maehr et al.”

“Specifically, the signals and conditions that underlie the differentiation of embryonic stem cells into PDX-1–positive pancreatic progenitor cells are now well established (36,37).”

“Human embryonic stem cells (hESCs) have an unlimited capacity for self-renewal and can differentiate into most or all cell types that belong to the ectoderm (4), mesoderm (5), and endoderm (6).”