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Oct4 is expressed in Nestin-positive cells as a marker for pancreatic endocrine progenitor

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Abstract

There are abundant progenitor cells in the developing pancreas, but molecular markers for these cells are lacking. Octamer-binding transcription factor-4 (Oct4) is an important transcription factor for keeping the features of self-renewal and pluripotency of embryonic stem cells. It’s well known that Oct4, as a totipotent stem cells marker, just is expressed in totipotent stem cells. In the present study, we collected ten human fetal pancreases, and found that Oct4 mRNA and protein were expressed in human fetal pancreas samples by RT-PCR, western blot and immunohistochemistry assays. Using double-staining, we demonstrated that Oct4 was not co-expressed with Chromogranin A (a peptide expressed in endocrine cells), but partially co-expressed with Ngn3 (a transcription factor expressed in pancreatic endocrine precursor cells) and Nestin (a intermediate filament, Nestin-positive cells isolated from islets can be induced to express insulin) in human fetal pancreases. Indeed, we prepared Nestin-positive cells from human fetal pancreas by cell selection, and found that these cells expressed Oct4 and Ngn3. The Nestin-positive cells displayed a rapid duplication and could differentiate into osteoblasts, fat and endocrine cells in vitro. These results indicated that the Nestin-positive cells in the fetal age should be pancreatic progenitor cells. Overall, our study suggested that Oct4 was a marker for pancreatic endocrine progenitor.

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Abbreviations

Oct4:

Octamer-binding transcription factor-4

POU5F1:

POU domain, class 5, transcription factor 1

POU:

Pit-Oct-Unc

Oct3:

Octamer-binding transcription factor-3

Ngn3:

Neurogenin 3

ESCs:

Embroynic stem cells

CgA:

Chromogranin A

DAPI:

4′,6-diamidino-2-phenylindole

bFGF:

Basic fibroblast growth factor

IBMX:

Methylisobutylxanthine

PFA:

Paraformaldehyde

CMF-PBS:

Ca2+and Mg2+ free PBS

ICCs:

Islet-like cell clusters

ALP:

Alkaline phosphatase

DTZ:

Dithizone

alpha-SMA:

Alpha-smooth muscle actin

qRT-PCR:

Quantitative real-time reverse transcription polymerase chain reaction

References

  • Apelqvist A, Li H, Sommer L, Beatus P, Anderson DJ, Honjo T, Hrabe de Angelis M, Lendahl U, Edlund H (1999) Notch signalling controls pancreatic cell differentiation. Nature 400:877–881

    Article  PubMed  CAS  Google Scholar 

  • Baertschiger RM, Bosco D, Morel P, Serre-Beinier V, Berney T, Buhler LH, Gonelle-Gispert C (2008) Mesenchymal stem cells derived from human exocrine pancreas express transcription factors implicated in beta-cell development. Pancreas 37:75–84

    Article  PubMed  CAS  Google Scholar 

  • Bernardo A, Barrow J, Hay C, McCreath K, Kind A, Schnieke A, Colman A, Hart A, Docherty K (2006) Presence of endocrine and exocrine markers in EGFP-positive cells from the developing pancreas of a Nestin/EGFP mouse. Mol Cell Endocrinol 253:14–21

    Article  PubMed  CAS  Google Scholar 

  • Blyszczuk P, Czyz J, Kania G, Wagner M, Roll U, St-Onge L, Wobus AM (2003) Expression of Pax4 in embryonic stem cells promotes differentiation of Nestin-positive progenitor and insulin-producing cells. Proc Natl Acad Sci USA 100:998–1003

    Article  PubMed  CAS  Google Scholar 

  • Collombat P, Hecksher-Sorensen J, Serup P, Mansouri A (2006) Specifying pancreatic endocrine cell fates. Mech Dev 123:501–512

    Article  PubMed  CAS  Google Scholar 

  • Delacour A, Nepote V, Trumpp A, Herrera P (2004) Nestin expression in pancreatic exocrine cell lineages. Mech Dev 121:3–14

    Article  PubMed  CAS  Google Scholar 

  • Esni F, Stoffers D, Takeuchi T, Leach S (2004) Origin of exocrine pancreatic cells from Nestin-positive precursors in developing mouse pancreas. Mech Dev 121:15–25

    Article  PubMed  CAS  Google Scholar 

  • Gasa R, Mrejen C, Leachman N, Otten M, Barnes M, Wang J, Chakrabarti S, Mirmira R, German M (2004) Proendocrine genes coordinate the pancreatic islet differentiation program in vitro. Proc Natl Acad Sci USA 101:13245–13250

    Article  PubMed  CAS  Google Scholar 

  • Gradwohl G, Dierich A, LeMeur M, Guillemot F (2000) Neurogenin3 is required for the development of the four endocrine cell lineages of the pancreas. Proc Natl Acad Sci USA 97:1607–1611

    Article  PubMed  CAS  Google Scholar 

  • Huang H, Tang X (2003) Phenotypic determination and characterization of Nestin-positive precursors derived from human fetal pancreas. Lab Invest 83:539–547

    PubMed  CAS  Google Scholar 

  • Jensen J, Heller RS, Funder-Nielsen T, Pedersen EE, Lindsell C, Weinmaster G, Madsen OD, Serup P (2000) Independent development of pancreatic alpha- and beta-cells from neurogenin3-expressing precursors: a role for the notch pathway in repression of premature differentiation. Diabetes 49:163–176

    Article  PubMed  CAS  Google Scholar 

  • Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, Reyes M, Lenvik T, Lund T, Blackstad M, Du J, Aldrich S, Lisberg A, Low WC, Largaespada DA, Verfaillie CM (2002) Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 418:41–49

    Article  PubMed  CAS  Google Scholar 

  • Lardon J, Rooman I, Bouwens L (2002) Nestin expression in pancreatic stellate cells and angiogenic endothelial cells. Histochem Cell Biol 117:535–540

    Article  PubMed  CAS  Google Scholar 

  • Lendahl U, Zimmerman LB, McKay RD (1990) CNS stem cells express a new class of intermediate filament protein. Cell 60:585–595

    Article  PubMed  CAS  Google Scholar 

  • 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 292:1389–1394

    Article  PubMed  CAS  Google Scholar 

  • Lyttle BM, Li J, Krishnamurthy M, Fellows F, Wheeler MB, Goodyer CG, Wang R (2008) Transcription factor expression in the developing human fetal endocrine pancreas. Diabetologia 51:1169–1180

    Article  PubMed  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C (T)) method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  • McGuckin CP, Forraz N, Baradez MO, Navran S, Zhao J, Urban R, Tilton R, Denner L (2005) Production of stem cells with embryonic characteristics from human umbilical cord blood. Cell Prolif 38:245–255

    Article  PubMed  CAS  Google Scholar 

  • Mish FC (1988) Webster s ninth new collegiate dictionary. Merriam-Webster, Markham

    Google Scholar 

  • Petri A, Ahnfelt-Ronne J, Frederiksen KS, Edwards DG, Madsen D, Serup P, Fleckner J, Heller RS (2006) The effect of neurogenin3 deficiency on pancreatic gene expression in embryonic mice. J Mol Endocrinol 37:301–316

    Article  PubMed  CAS  Google Scholar 

  • Sander M, German MS (1997) The beta cells transcription factors and development of the pancreas. J Mol Med 75:327–340

    Article  PubMed  CAS  Google Scholar 

  • Scholer HR, Ruppert S, Suzuki N, Chowdhury K, Gruss P (1990) New type of POU domain in germ line-specific protein Oct-4. Nature 344:435–439

    Article  PubMed  CAS  Google Scholar 

  • Schwitzgebel VM, Scheel DW, Conners JR, Kalamaras J, Lee JE, Anderson DJ, Sussel L, Johnson JD, German MS (2000) Expression of neurogenin3 reveals an islet cell precursor population in the pancreas. Development 127:3533–3542

    PubMed  CAS  Google Scholar 

  • Selander L, Edlund H (2002) Nestin is expressed in mesenchymal and not epithelial cells of the developing mouse pancreas. Mech Dev 113:189–192

    Article  PubMed  CAS  Google Scholar 

  • Serafimidis I, Rakatzi I, Episkopou V, Gouti M, Gavalas A (2008) Novel effectors of directed and Ngn3-mediated differentiation of mouse embryonic stem cells into endocrine pancreas progenitors. Stem Cells 26:3–16

    Article  PubMed  CAS  Google Scholar 

  • Sommer L, Ma Q, Anderson DJ (1996) Neurogenins, a novel family of atonal-related bHLH transcription factors, are putative mammalian neuronal determination genes that reveal progenitor cell heterogeneity in the developing CNS and PNS. Mol Cell Neurosci 8:221–241

    Article  PubMed  CAS  Google Scholar 

  • Treutelaar M, Skidmore J, Dias-Leme C, Hara M, Zhang L, Simeone D, Martin D, Burant C (2003) Nestin-lineage cells contribute to the microvasculature but not endocrine cells of the islet. Diabetes 52:2503–2512

    Article  PubMed  CAS  Google Scholar 

  • Wang R, Li J, Yashpal N (2004) Phenotypic analysis of c-Kit expression in epithelial monolayers derived from postnatal rat pancreatic islets. J Endocrinol 182:113–122

    Article  PubMed  CAS  Google Scholar 

  • Wilson M, Scheel D, German M (2003) Gene expression cascades in pancreatic development. Mech Dev 120:65–80

    Article  PubMed  CAS  Google Scholar 

  • Yashpal N, Li J, Wang R (2004) Characterization of c-Kit and Nestin expression during islet cell development in the prenatal and postnatal rat pancreas. Dev Dyn 229:813–825

    Article  PubMed  CAS  Google Scholar 

  • Zhang YQ, Kritzik M, Sarvetnick N (2005) Identification and expansion of pancreatic stem/progenitor cells. J Cell Mol Med 9:331–344

    Article  PubMed  CAS  Google Scholar 

  • Zhao M, Amiel SA, Christie MR, Muiesan P, Srinivasan P, Littlejohn W, Rela M, Arno M, Heaton N, Huang GC (2007) Evidence for the presence of stem cell-like progenitor cells in human adult pancreas. J Endocrinol 195:407–414

    Article  PubMed  CAS  Google Scholar 

  • Zhao M, Amiel SA, Ajami S, Jiang J, Rela M, Heaton N, Huang GC (2008) Amelioration of streptozotocin-induced diabetes in mice with cells derived from human marrow stromal cells. PLoS ONE 3:e2666

    Article  PubMed  CAS  Google Scholar 

  • Zulewski H, Abraham EJ, Gerlach MJ, Daniel PB, Moritz W, Muller B, Vallejo M, Thomas MK, Habener JF (2001) Multipotential Nestin-positive stem cells isolated from adult pancreatic islets differentiate ex vivo into pancreatic endocrine, exocrine, and hepatic phenotypes. Diabetes 50:521–533

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Vincent Wong, Changyong Li for paper writing.

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Authors and Affiliations

  1. Stem Cell Research Center, Health Science Center, Peking University, No. 38 Xueyuan Road, Haidian District, Beijing, China

    Hong Wang, Shuling Wang, Jiang Hu, Yaxian Kong, Shen Chen & Lingsong Li

  2. Department of Cell Biology, Capital Medical University, No. 10 Xitoutiao, You An Men, Beijing, China

    Liying Li

Authors
  1. Hong Wang
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  2. Shuling Wang
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  3. Jiang Hu
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  4. Yaxian Kong
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  5. Shen Chen
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  6. Lingsong Li
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  7. Liying Li
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Corresponding author

Correspondence to Liying Li.

Additional information

H. Wang and S. Wang contributed equally to this work.

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Wang, H., Wang, S., Hu, J. et al. Oct4 is expressed in Nestin-positive cells as a marker for pancreatic endocrine progenitor. Histochem Cell Biol 131, 553–563 (2009). https://doi.org/10.1007/s00418-009-0560-x

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