Skip to main content

Advertisement

SpringerLink
Log in

Migration and differentiation of transplanted enteric neural crest-derived cells in murine model of Hirschsprung’s disease

  • JAACT Special Issue
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

Stem cell therapy offers the potential of rebuilding the enteric nervous system (ENS) in the aganglionic bowel of patients with Hirschsprung’s disease. P0-Cre/Floxed-EGFP mice in which neural crest-derived cells express EGFP were used to obtain ENS stem/progenitor cells. ENS stem/progenitor cells were transplanted into the bowel of Ret−/− mouse, an animal model of Hirschsprung’s disease. Immunohistochemical analysis was performed to determine whether grafted cells gave rise to neurons in the recipient bowel. EGFP expressing neural crest-derived cells accounted for 7.01 ± 2.52 % of total cells of gastrointestinal tract. ENS stem/progenitor cells were isolated using flow cytometry and expanded as neurosphere-like bodies (NLBs) in a serum-free culture condition. Some cells in NLBs expressed neural crest markers, p75 and Sox10 and neural stem/progenitor cells markers, Nestin and Musashi1. Multipotency of isolated ENS stem/progenitor cells was determined as they differentiated into neurons, glial cells, and myofibloblasts in culture. When co-cultured with explants of hindgut of Ret−/− mice, ENS stem/progenitor cells migrated into the aganglionic bowel and gave rise to neurons. ENS stem/progenitor cells used in this study appear to be clinically relevant donor cells in cell therapy to treat Hirschsprung’s disease capable of colonizing the affected bowel and giving rise to neurons.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Almond S, Lindley RM, Kenny SE, Connell MG, Edgar DH (2007) Characterisation and transplantation of enteric nervous system progenitor cells. Gut 56:489–496

    Article  Google Scholar 

  • Azan G, Low WC, Wendelschafer-Crabb G, Ikramuddin S, Kennedy WR (2011) Evidence for neural progenitor cells in the human adult enteric nervous system. Cell Tissue Res 344:217–225

    Article  Google Scholar 

  • Becker L, Peterson J, Kulkarni S, Pasricha PJ (2013) Ex vivo neurogenesis within enteric ganglia occurs in a PTEN dependent manner. PLoS One 8:e59452

    Article  CAS  Google Scholar 

  • Belkind-Gerson J, Carreon-Rodriguez A, Benedict LA, Steiger C, Pieretti A, Nagy N, Dietrich J, Goldstein AM (2013) Nestin-expressing cells in the gut give rise to enteric neurons and glial cells. Neurogastroenterol Motil 25:61–69

    Article  CAS  Google Scholar 

  • Bondurand N, Natarajan D, Thapar N, Atkins C, Pachnis V (2003) Neuron and glia generating progenitors of the mammalian enteric nervous system isolated from foetal and postnatal gut cultures. Development 130:6387–6400

    Article  CAS  Google Scholar 

  • Bondurand N, Natarajan D, Barlow A, Thapar N, Pachnis V (2006) Maintenance of mammalian enteric nervous system progenitors by SOX10 and endothelin 3 signalling. Development 133:2075–2086

    Article  CAS  Google Scholar 

  • Breau MA, Pietri T, Eder O, Blanche M, Brakebusch C, Fässler R, Thiery JP, Dufour S (2006) Lack of beta1 integrins in enteric neural crest cells leads to a Hirschsprung-like phenotype. Development 133:1725–1734

    Article  CAS  Google Scholar 

  • Catto-Smith AG, Trajanovska M, Taylor RG (2007) Long-term continence after surgery for Hirschsprung’s disease. J Gastroenterol Hepatol 22:2273–2282

    Article  Google Scholar 

  • Durbec PL, Larsson-Blomberg LB, Schuchardt A, Costantini F, Pachnis V (1996) Common origin and developmental dependence on c-ret of subsets of enteric and sympathetic neuroblasts. Development 122:349–358

    CAS  Google Scholar 

  • Fu M, Vohra BP, Wind D, Heuckeroth RO (2006) BMP signaling regulates murine enteric nervous system precursor migration, neurite fasciculation, and patterning via altered Ncam1 polysialic acid addition. Dev Biol 299:137–150

    Article  CAS  Google Scholar 

  • Furness JB (2012) The enteric nervous system and neurogastroenterology. Nat Rev Gastroenterol Hepatol 9:286–294

    Article  CAS  Google Scholar 

  • Goldstein AM, Brewer KC, Doyle AM, Nagy N, Roberts DJ (2005) BMP signaling is necessary for neural crest cell migration and ganglion formation in the enteric nervous system. Mech Dev 122:821–833

    Article  CAS  Google Scholar 

  • Hearn CJ, Young HM, Ciampoli D, Lomax AE, Newgreen D (1999) Catenary cultures of embryonic gastrointestinal tract support organ morphogenesis, motility, neural crest cell migration, and cell differentiation. Dev Dyn 214:239–247

    Article  CAS  Google Scholar 

  • Hotta R, Stamp L, Foong JP, McConnell SN, Bergner AJ, Anderson RB, Enomoto H, Newgreen DF, Obermayr F, Furness JB, Young HM (2013) Transplanted progenitors generate functional enteric neurons in the postnatal colon. J Clin Invest 123:1182–1191

    Article  CAS  Google Scholar 

  • Ieiri S, Nakatsuji T, Akiyoshi J, Higashi M, Hashizume M, Suita S, Taguchi T (2010) Long-term outcomes and the quality of life of Hirschsprung disease in adolescents who have reached 18 years or older-a 47-year single-institute experience. J Pediatr Surg 45:2398–2402

    Article  Google Scholar 

  • Joseph NM, He S, Quintana E, Kim YG, Núñez G, Morrison SJ (2011) Enteric glia are multipotent in culture but primarily form glia in the adult rodent gut. J Clin Invest 121:3398–3411

    Article  CAS  Google Scholar 

  • Kaneko Y, Sakakibara S, Imai T, Suzuki A, Nakamura Y, Sawamoto K, Ogawa Y, Toyama Y, Miyata T, Okano H (2000) Musashi1: an evolutionally conserved marker for CNS progenitor cells including neural stem cells. Dev Neurosci 22:139–153

  • Kawamoto S, Niwa H, Tashiro F, Sano S, Kondoh G, Takeda J, Tabayashi K, Miyazaki J (2000) A novel reporter mouse strain that expresses enhanced green fluorescent protein upon Cre-mediated recombination. FEBS Lett 470:263–268

  • Kruger G, Mosher J, Bixby S, Joseph N, Iwashita T, Morrison SJ (2002) Neural crest stem cells persist in the adult gut but undergo changes in self-renewal, neuronal subtype potential, and factor responsiveness. Neuron 35:657–669

  • Laranjeira C, Sandgren K, Kessaris N, Richardson W, Potocnik A, Vanden Berghe P, Pachnis V (2011) Glial cells in the mouse enteric nervous system can undergo neurogenesis in response to injury. J Clin Invest 121:3412–3424

  • Lee MJ, Calle E, Brennan A, Ahmed S, Sviderskaya E, Jessen KR, Mirsky R (2001) In early development of the rat mRNA for the major myelin protein P(0) is expressed in nonsensory areas of the embryonic inner ear, notochord, enteric nervous system, and olfactory ensheathing cells. Dev Dyn 222:40–51

    Article  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  CAS  Google Scholar 

  • Lindley RM, Hawcutt DB, Connell MG, Almond SL, Vannucchi MG, Faussone-Pellegrini MS, Edgar DH, Kenny SE (2008) Human and mouse enteric nervous system neurosphere transplants regulate the function of aganglionic embryonic distal colon. Gastroenterology 135:205–216

  • Liu MT, Kuan YH, Wang J et al (2009) 5-HT4 receptor-mediated neuroprotection and neurogenesis in the enteric nervous system of adult mice. J Neurosci 29:9683-9699

    Article  CAS  Google Scholar 

  • Matsuzaki Y, Kinjo K, Mulligan RC, Okano H (2004) Unexpectedly efficient homing capacity of purified murine hematopoietic stem cells. Immunity 20:87–93

  • Metzger M, Caldwell C, Barlow AJ, Burns AJ, Thapar N (2009) Enteric nervous system stem cells derived from human gut mucosa for the treatment of aganglionic gut disorders. Gastroenterology 136:2214–2225

    Article  CAS  Google Scholar 

  • Mosher JT, Yeager KJ, Kruger GM, Joseph NM, Hutchin ME, Dlugosz AA, Morrison SJ (2007) Intrinsic differences among spatially distinct neural crest stem cells in terms of migratory properties, fate determination, and ability to colonize the enteric nervous system. Dev Biol 303:1–15

  • Nagoshi N, Shibata S, Kubota Y, Nakamura M, Nagai Y, Satoh E, Morikawa S, Okada Y, Mabuchi Y, Katoh H, Okada S, Fukuda K, Suda T, Matsuzaki Y, Toyama Y, Okano H (2008) Ontogeny and multipotency of neural crest-derived stem cells in mouse bone marrow, dorsal root ganglia, and whisker pad. Cell Stem Cell 2:392–403

  • Natarajan D, Grigoriou M, Marcos-Gutierrez CV, Atkins C, Pachnis V (1999) Multipotential progenitors of the mammalian enteric nervous system capable of colonising aganglionic bowel in organ culture. Development 126:157–168

    CAS  Google Scholar 

  • Okano H, Imai T, Okabe M (2002) Musashi: a translational regulator of cell fate. J Cell Sci 115:1355–1359

    CAS  Google Scholar 

  • Paratore C, Goerich DE, Suter U, Wegner M, Sommer L (2001) Survival and glial fate acquisition of neural crest cells are regulated by an interplay between the transcription factor Sox10 and extrinsic combinatorial signaling. Development 128:3949–3961

  • Potten CS, Booth C, Tudor GL, Booth D, Brady G, Hurley P, Ashton G, Clarke R, Sakakibara S, Okano H (2003) Identification of a putative intestinal stem cell and early lineage marker; musashi-1. Differentiation 71:28–41

  • Reynolds BA, Weiss S (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255:1707–1710

    Article  CAS  Google Scholar 

  • Sakakibara S, Imai T, Hamaguchi K, Okabe M, Aruga J, Nakajima K, Yasutomi D, Nagata T, Kurihara Y, Uesugi S, Miyata T, Ogawa M, Mikoshiba K, Okano H (1996) Mouse-Musashi-1, a neural RNA-binding protein highly enriched in the mammalian CNS stem cell. Dev Biol 176:230–242

  • Sawamoto K, Nakao N, Kobayashi K, Matsushita N, Takahashi H, Kakishita K, Yamamoto A, Yoshizaki T, Terashima T, Murakami F, Itakura T, Okano H (2001) Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons. Proc Natl Acad Sci USA 98:6423–6428

  • Schuchardt A, D’Agati V, Larsson-Blomberg L, Costantini F, Pachnis V (1994) Defects in the kidney and enteric nervous system of mice lacking the tyrosine kinase receptor Ret. Nature 367:380–383

  • Stemple DL, Anderson DJ (1992) Isolation of a stem cell for neurons and glia from the mammalian neural crest. Cell 71:973–985

    Article  CAS  Google Scholar 

  • Suarez-Rodriguez R, Belkind-Gerson J (2004) Cultured nestin-positive cells from postnatal mouse small bowel differentiate ex vivo into neurons, glia, and smooth muscle. Stem Cells 22:1373–1385

    Article  Google Scholar 

  • Sun NF, Zhong WY, Lu SA, Tian YL, Chen JB, Hu SY, Tian AL (2013) Coexpression of recombinant adenovirus carrying GDNF and EDNRB genes in neural stem cells in vitro. Cell Biol Int 37:458–463

  • Tomita Y, Matsumura K, Wakamatsu Y, Matsuzaki Y, Shibuya I, Kawaguchi H, Ieda M, Kanakubo S, Shimazaki T, Ogawa S, Osumi N, Okano H, Fukuda K (2005) Cardiac neural crest cells contribute to the dormant multipotent stem cell in the mammalian heart. J Cell Biol 170:1135–1146

  • Yamauchi Y, Abe K, Mantani A, Hitoshi Y, Suzuki M, Osuzu F, Kuratani S, Yamamura K (1999) A novel transgenic technique that allows specific marking of the neural crest cell lineage in mice. Dev Biol 212:191–203

  • Yoshida S, Shimmura S, Nagoshi N, Fukuda K, Matsuzaki Y, Okano H, Tsubota K (2006) Isolation of multipotent neural crest-derived stem cells from the adult mouse cornea. Stem Cells 24:2714–2722

  • Young HM, Bergner AJ, Anderson RB, Enomoto H, Milbrandt J, Newgreen DF, Whitington PM (2004) Dynamics of neural crest-derived cell migration in the embryonic mouse gut. Dev Biol 270:455–473

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank M Mori, T Harada, S Suzuki (FACS technician) for technical supports and animal care, M Jijiwa for information about Ret−/− mice, HM Young for helpful discussion to prepare manuscript, M Takahashi for providing Ret−/− mice, F Costantini for giving a permission to have a Ret−/− mice. This work was supported by the Ministry of Education, Science, and Culture of Japan (MEXT), KAKENHI, Grant-in-Aid for Scientific Research (C) 20592091 and Grant-in-Aid for Young Scientists (B) 21791736, by a Grant-in-aid from the Global COE program of MEXT to Keio University.

Author information

Author notes

  1. Ryuhei Nishikawa

    Present address: Department of Dermatology, Kurume University School of Medicine, Kurume University Institute of Cutaneous Cell Biology, 67 Asahimachi, Kurume, Fukuoka, 830-0011, Japan

  2. Ryo Hotta

    Present address: Department of Pediatric Surgery, Massachusetts General Hospital and Harvard Medical School, 55 Fruit Street Warren 1153, Boston, MA, 02114, USA

  3. Naoki Shimojima

    Present address: Department of Surgery, Tokyo Metropolitan Children’s Medical Center, 2-8-29 Musashidai, Fuchu-city, Tokyo, 183-8561, Japan

  4. Yumi Matsuzaki

    Present address: Institute of Integral Medical Research, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan

  5. Hirotaka J. Okano

    Present address: Division of Regenerative Medicine, Jikei University School of Medicine, 3-25-8 Nishi-Shinbashi, Minato-ku, Tokyo, 105-0003, Japan

  6. Yasuhide Morikawa

    Present address: Department of Pediatric Surgery, International University of Health and Welfare, 537-3 Iguchi, Nasushiobara, Tochigi, 329-2763, Japan

Authors and Affiliations

  1. Department of Pediatric Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan

    Ryuhei Nishikawa, Ryo Hotta, Naoki Shimojima, Tatsuo Kuroda & Yasuhide Morikawa

  2. Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan

    Shinsuke Shibata, Yumi Matsuzaki, Hirotaka J. Okano & Hideyuki Okano

  3. Department of Orthopedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan

    Narihito Nagoshi & Masaya Nakamura

Authors
  1. Ryuhei Nishikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ryo Hotta
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Naoki Shimojima
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shinsuke Shibata
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Narihito Nagoshi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Masaya Nakamura
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yumi Matsuzaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hirotaka J. Okano
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Tatsuo Kuroda
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Hideyuki Okano
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Yasuhide Morikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Naoki Shimojima.

Additional information

Ryuhei Nishikawa and Ryo Hotta equally contributed to this work.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nishikawa, R., Hotta, R., Shimojima, N. et al. Migration and differentiation of transplanted enteric neural crest-derived cells in murine model of Hirschsprung’s disease. Cytotechnology 67, 661–670 (2015). https://doi.org/10.1007/s10616-014-9754-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-014-9754-8

Keywords

Search

Navigation