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Cytotechnology

, Volume 68, Issue 4, pp 987–998 | Cite as

Development of an optimized 5-stage protocol for the in vitro preparation of insulin-secreting cells from mouse ES cells

  • Mikako SaitoEmail author
  • Asako Kaneda
  • Hajime Shigeto
  • Nobuaki Hanata
  • Keiko Otokuni
  • Hideaki Matsuoka
Original Research

Abstract

In order to produce insulin-secreting cells with a high value of glucose-stimulated insulin secretion (GSIS) from mouse embryonic stem cells, we have developed an optimized 5-stage protocol by referring to culture conditions so far reported elsewhere. This protocol is characterized by 4 points: (1) use of an activin-free medium in the first stage, (2) use of gelatin/fibronectin coated culture dishes in 1–4 stages throughout, (3) removal of undifferentiated cells by cell sorter at the end of 4th stage, and (4) sedimental culture in the 5th stage. GSIS value of the produced cells reached 2.4, that was at a higher rank of those so far reported. The produced cells were transplanted in diabetes model mice but no remedy effect was observed. Then transplantation was conducted in pre-diabetes model mice, in which GSIS was impaired without affecting insulin producing function. The transplantation of 5 × 106 cells resulted in a marked improvement of glucose tolerance within 20 days. This effect decreased but was still observed at 120 days post-transplantation. This demonstrates the feasibility of the novel optimized protocol.

Keywords

Glucose-stimulated insulin secretion Insulin-secreting cells Mouse ES cells Pre-diabetes model mice 

Notes

Acknowledgments

We thank Dr. Niwa for the donation of feeder free EB3 cells, and Dr. Okitsu for excellent technical support on transplantation. The work was supported in part by the Strategic Research Promotion Program, the Ministry of Education, Culture, Sports, Science, and Technology, on the research subject “Development of Next Generation Bioresources”.

References

  1. Beattie GM, Lopez AD, Bucay N, Hinton A, Firpo MT, King CC, Hayek A (2005) Activin A maintains pluripotency of human embryonic stem cells in the absence of feeder layers. Stem Cells 23:489–495CrossRefGoogle Scholar
  2. D’Amour KA, Bang AG, Eliazer S, Kelly OG, Agulnick AD, Smart NG, Moorman MA, Kroon E, Carpenter MK, Baetge EE (2006) Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells. Nat Biotechnol 24:1392–1401CrossRefGoogle Scholar
  3. James D, Levine AJ, Besser D, Hemmati-Brivanlou A (2005) TGFbeta/activin/nodal signaling is necessary for the maintenance of pluripotency in human embryonic stem cells. Development 132:1273–1282CrossRefGoogle Scholar
  4. Jiang W, Shi Y, Zhao D, Chen S, Yong J, Zhang J, Qing T, Sun X, Zhang P, Ding M, Li D, Deng H (2007) In vitro derivation of functional insulin-producing cells from human embryonic stem cells. Cell Res 17:333–344CrossRefGoogle Scholar
  5. Komatsu M, Takei M, Ishii H, Sato Y (2013) Glucose-stimulated insulin secretion: a newer perspective. J Diabetes Investig 4:511–516CrossRefGoogle Scholar
  6. Kroon E, Martinson LA, Kadoya K, Bang AG, Kelly OG, Eliazer S, Young H, Richardson M, Smart NG, Cunningham J, Agulnick AD, D’Amour KA, Carpenter MK, Baetge EE (2008) Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo. Nat Biotechnol 26:443–452CrossRefGoogle Scholar
  7. Kubo A, Shinozaki K, Shannon JM, Kouskoff V, Kennedy M, Woo S, Fehling HJ, Keller G (2004) Development of definitive endoderm from embryonic stem cells in culture. Development 131:1651–1662CrossRefGoogle Scholar
  8. Kuliawat R, Klein L, Gong Z, Nicoletta-Gentile M, Nemkal A, Cui L, Bastie C, Su K, Huffman D, Surana M, Barzilai N, Fleischer N, Muzumdar R (2013) Potent humanin analog increases glucose-stimulated insulin secretion through enhanced metabolism in the cell. FASEB J 27:4890–4898CrossRefGoogle Scholar
  9. Kunisada Y, Tsubooka-Yamazoe N, Shoji M, Hosoya M (2012) Small molecules induce efficient differentiation into insulin-producing cells from human induced pluripotent stem cells. Stem Cell Res 8:274–284CrossRefGoogle Scholar
  10. Lumelsky N, Blondel O, Laeng P, Velasco I, Ravin R, McKay R (2001) Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets. Science 292:1389–1394CrossRefGoogle Scholar
  11. Moritoh Y, Yamato E, Yasui Y, Miyazaki S, Miyazaki J (2003) Analysis of insulin-producing cells during in vitro differentiation from feeder-free embryonic stem cells. Diabetes 52:1163–1168CrossRefGoogle Scholar
  12. Ogawa K, Saito A, Matsui H, Suzuki H, Ohtsuka S, Shimosato D, Morishita Y, Watabe T, Niwa H, Miyazono K (2007) Activin-Nodal signaling is involved in propagation of mouse embryonic stem cells. J Cell Sci 120:55–65CrossRefGoogle Scholar
  13. Ohara-Imaizumi M, Kim H, Yoshida M, Fujiwara T, Aoyagi K, Toyofuku Y, Nakamichi Y, Nishiwaki C, Okamura T, Uchida T, Fujitani Y, Akagawa K, Kakei M, Watada H, German MS, Nagamatsu S (2013) Serotonin regulates glucose-stimulated insulin secretion from pancreatic β cells during pregnancy. Proc Natl Acad Sci USA 110:19420–19425CrossRefGoogle Scholar
  14. Parashurama N, Nahmias Y, Cho CH, van Poll D, Tilles AW, Berthiaume F, Yarmush ML (2008) Activin alters the kinetics of endoderm induction in embryonic stem cells cultured on collagen gels. Stem Cells 26:474–484CrossRefGoogle Scholar
  15. Postic C, Shiota M, Niswender KD, Jetton TL, Chen Y, Moates JM, Shelton KD, Lindner J, Cherrington AD, Magnuson MA (1999) Dual roles for glucokinase in glucose homeostasis as determined by liver and pancreatic β cell-specific gene knock-outs using cre recombinase. J Biol Chem 274:305–315CrossRefGoogle Scholar
  16. Pound LD, Oeser JK, O’Brien TP, Wang Y, Faulman CJ, Dadi PK, Jacobson DA, Hutton JC, McGuinness OP, Shiota M, O’Brien RM (2013) G6PC2: a negative regulator of basal glucose-stimulated insulin secretion. Diabetes 62:1547–1556CrossRefGoogle Scholar
  17. Printz RL, Granner DK (2005) Tweaking the glucose sensor: adjusting glucokinase activity with activator compounds. Endocrinology 146:3693–3695CrossRefGoogle Scholar
  18. Saito M, Hayakawa A, Inagaki N, Matsuoka H (2013) Development of novel cell lines of diabetic dysfunction model fit for cell-based screening tests of medicinal materials. Cytotechnology 65:105–118CrossRefGoogle Scholar
  19. Sakano D, Shiraki N, Kikawa K, Yamazoe T, Kataoka M, Umeda K, Araki K, Mao D, Matsumoto S, Nakagata N, Andersson O, Stainier D, Endo F, Kume K, Uesugi M, Kume S (2014) VMAT2 identified as a regulator of late-stage β-cell differentiation. Nat Chem Biol 10:141–148CrossRefGoogle Scholar
  20. Schäfer MK, Hartwig NR, Kalmbach N, Klietz M, Anlauf M, Eiden LE, Weihe E (2013) Species-specific vesicular monoamine transporter 2 (VMAT2) expression in mammalian pancreatic beta cells: implications for optimising radioligand-based human beta cell mass (BCM) imaging in animal models. Diabetologia 56:1047–1056CrossRefGoogle Scholar
  21. Sherwood RI, Jitianu C, Cleaver O, Shaywitz DA, Lamenzo JO, Chen AE, Golub TR, Melton DA (2007) Prospective isolation and global gene expression analysis of definitive and visceral endoderm. Dev Biol 304:541–555CrossRefGoogle Scholar
  22. Soria B (2001) In-vitro differentiation of pancreatic beta-cells. Differentiation 68:205–219CrossRefGoogle Scholar
  23. Stainier DY (2002) A glimpse into the molecular entrails of endoderm formation. Genes Dev 16:893–907CrossRefGoogle Scholar
  24. Takeuchi H, Nakatsuji N, Suemori H (2014) Endodermal differentiation of human pluripotent stem cells to insulin-producing cells in 3D culture. Sci Rep 4:4488. doi: 10.1038/srep04488 Google Scholar
  25. Tuch BE, Gao SY, Lees JG (2014) Scaffolds for islets and stem cells differentiated into insulin-secreting cells. Front. Biosci. 19:126–138CrossRefGoogle Scholar
  26. Vallier L, Alexander M, Pedersen RA (2005) Activin/Nodal and FGF pathways cooperate to maintain pluripotency of human embryonic stem cells. J Cell Sci 118:4495–4509CrossRefGoogle Scholar
  27. Veras K, Almeida FN, Nachbar RT, de Jesus DS, Camporez JP, Carpinelli AR, Goedecke JH, de Oliveira Carvalho CR (2014) DHEA supplementation in ovariectomized rats reduces impaired glucose-stimulated insulin secretion induced by a high-fat diet. FEBS Open Bio 4:141–146CrossRefGoogle Scholar
  28. Yasunaga M, Tada S, Torikai-Nishikawa S, Nakano Y, Okada M, Jakt LM, Nishikawa S, Chiba T, Era T, Nishikawa S (2005) Induction and monitoring of definitive and visceral endoderm differentiation of mouse ES cells. Nat Biotechnol 23:1542–1550CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Mikako Saito
    • 1
    Email author
  • Asako Kaneda
    • 1
  • Hajime Shigeto
    • 1
  • Nobuaki Hanata
    • 1
  • Keiko Otokuni
    • 1
  • Hideaki Matsuoka
    • 1
  1. 1.Department of Biotechnology and Life ScienceTokyo University of Agriculture and TechnologyKoganeiJapan

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