Development of an Optimized 5-stage Protocol for the in Vitro Preparation of Insulin-secreting Cells from Mouse ES Cells

Overview

Journal Cytotechnology

Specialties Biotechnology
Genetics

Date 2015 Mar 10

PMID 25749915

Citations 5

Authors

Mikako Saito

Asako Kaneda

Hajime Shigeto

Nobuaki Hanata

Keiko Otokuni

Hideaki Matsuoka

Affiliations

Soon will be listed here.

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 × 10(6) 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.

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References

Lumelsky N, Blondel O, Laeng P, Velasco I, Ravin R, McKay R . Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets. Science. 2001; 292(5520):1389-94. DOI: 10.1126/science.1058866. View

Jiang W, Shi Y, Zhao D, Chen S, Yong J, Zhang J . In vitro derivation of functional insulin-producing cells from human embryonic stem cells. Cell Res. 2007; 17(4):333-44. DOI: 10.1038/cr.2007.28. View

Parashurama N, Nahmias Y, Cho C, van Poll D, Tilles A, Berthiaume F . Activin alters the kinetics of endoderm induction in embryonic stem cells cultured on collagen gels. Stem Cells. 2007; 26(2):474-84. PMC: 2802581. DOI: 10.1634/stemcells.2007-0303. View

James D, Levine A, Besser D, Hemmati-Brivanlou A . TGFbeta/activin/nodal signaling is necessary for the maintenance of pluripotency in human embryonic stem cells. Development. 2005; 132(6):1273-82. DOI: 10.1242/dev.01706. View

Veras K, Almeida F, Nachbar R, de Jesus D, Camporez J, Carpinelli A . DHEA supplementation in ovariectomized rats reduces impaired glucose-stimulated insulin secretion induced by a high-fat diet. FEBS Open Bio. 2014; 4:141-6. PMC: 3907747. DOI: 10.1016/j.fob.2014.01.005. View

Pound L, Oeser J, OBrien T, Wang Y, Faulman C, Dadi P . G6PC2: a negative regulator of basal glucose-stimulated insulin secretion. Diabetes. 2013; 62(5):1547-56. PMC: 3636628. DOI: 10.2337/db12-1067. View

Tuch B, Gao S, Lees J . Scaffolds for islets and stem cells differentiated into insulin-secreting cells. Front Biosci (Landmark Ed). 2014; 19(1):126-38. DOI: 10.2741/4199. View

Ohara-Imaizumi M, Kim H, Yoshida M, Fujiwara T, Aoyagi K, Toyofuku Y . Serotonin regulates glucose-stimulated insulin secretion from pancreatic β cells during pregnancy. Proc Natl Acad Sci U S A. 2013; 110(48):19420-5. PMC: 3845121. DOI: 10.1073/pnas.1310953110. View

DAmour K, Bang A, Eliazer S, Kelly O, Agulnick A, Smart N . Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells. Nat Biotechnol. 2006; 24(11):1392-401. DOI: 10.1038/nbt1259. View

10.

Sakano D, Shiraki N, Kikawa K, Yamazoe T, Kataoka M, Umeda K . VMAT2 identified as a regulator of late-stage β-cell differentiation. Nat Chem Biol. 2013; 10(2):141-8. DOI: 10.1038/nchembio.1410. View

11.

Saito M, Hayakawa A, Inagaki N, Matsuoka H . Development of novel cell lines of diabetic dysfunction model fit for cell-based screening tests of medicinal materials. Cytotechnology. 2012; 65(1):105-18. PMC: 3536874. DOI: 10.1007/s10616-012-9466-x. View

12.

Beattie G, Lopez A, Bucay N, Hinton A, Firpo M, King C . Activin A maintains pluripotency of human embryonic stem cells in the absence of feeder layers. Stem Cells. 2005; 23(4):489-95. DOI: 10.1634/stemcells.2004-0279. View

13.

Stainier D . A glimpse into the molecular entrails of endoderm formation. Genes Dev. 2002; 16(8):893-907. DOI: 10.1101/gad.974902. View

14.

Soria B . In-vitro differentiation of pancreatic beta-cells. Differentiation. 2002; 68(4-5):205-19. DOI: 10.1046/j.1432-0436.2001.680408.x. View

15.

Printz R, Granner D . Tweaking the glucose sensor: adjusting glucokinase activity with activator compounds. Endocrinology. 2005; 146(9):3693-5. DOI: 10.1210/en.2005-0689. View

16.

Kubo A, Shinozaki K, Shannon J, Kouskoff V, Kennedy M, Woo S . Development of definitive endoderm from embryonic stem cells in culture. Development. 2004; 131(7):1651-62. DOI: 10.1242/dev.01044. View

17.

Kunisada Y, Tsubooka-Yamazoe N, Shoji M, Hosoya M . Small molecules induce efficient differentiation into insulin-producing cells from human induced pluripotent stem cells. Stem Cell Res. 2011; 8(2):274-84. DOI: 10.1016/j.scr.2011.10.002. View

18.

Yasunaga M, Tada S, Torikai-Nishikawa S, Nakano Y, Okada M, Jakt L . Induction and monitoring of definitive and visceral endoderm differentiation of mouse ES cells. Nat Biotechnol. 2005; 23(12):1542-50. DOI: 10.1038/nbt1167. View

19.

Postic C, Shiota M, Niswender K, Jetton T, Chen Y, Moates J . Dual roles for glucokinase in glucose homeostasis as determined by liver and pancreatic beta cell-specific gene knock-outs using Cre recombinase. J Biol Chem. 1998; 274(1):305-15. DOI: 10.1074/jbc.274.1.305. View

20.

Moritoh Y, Yamato E, Yasui Y, Miyazaki S, Miyazaki J . Analysis of insulin-producing cells during in vitro differentiation from feeder-free embryonic stem cells. Diabetes. 2003; 52(5):1163-8. DOI: 10.2337/diabetes.52.5.1163. View