Optimizing neurogenic potential of enteric neurospheres for treatment of neurointestinal diseases

Lily S. Cheng, Hannah K. Graham, Wei Hua Pan, N. Nagy, Alfonso Carreon-Rodriguez, Allan M. Goldstein, Ryo Hotta

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Background Enteric neurospheres derived from postnatal intestine represent a promising avenue for cell replacement therapy to treat Hirschsprung disease and other neurointestinal diseases. We describe a simple method to improve the neuronal yield of spontaneously formed gut-derived neurospheres. Materials and methods Enteric neurospheres were formed from the small and large intestines of mouse and human subjects. Neurosphere size, neural crest cell content, cell migration, neuronal differentiation, and neuronal proliferation in culture were analyzed. The effect of supplemental neurotrophic factors, including glial cell line-derived neurotrophic factor (GDNF) and endothelin-3, was also assessed. Results Mouse small intestine–derived neurospheres contained significantly more P75-expressing neural crest-derived cells (49.9 ± 15.3% versus 21.6 ± 11.9%, P < 0.05) and gave rise to significantly more Tuj1-expressing neurons than colon-derived neurospheres (69.9 ± 8.6% versus 46.2 ± 15.6%, P < 0.05). A similar pattern was seen in neurospheres isolated from human small and large intestine (32.6 ± 17.5% versus 10.2 ± 8.2% neural crest cells, P < 0.05; 29.7 ± 16.4% versus 16.0 ± 13.5% enteric neurons, P < 0.05). The addition of GDNF to the culture media further improved the neurogenic potential of small intestinal neurospheres (75.9 ± 4.0% versus 67.8 ± 5.8%, P < 0.05) whereas endothelin-3 had no effect. Conclusions Enteric neurospheres formed from small intestine and supplemented with GDNF yield an enriched population of neural crest-derived progenitor cells and give rise to a high density of enteric neurons.

Original languageEnglish
Pages (from-to)451-459
Number of pages9
JournalJournal of Surgical Research
Volume206
Issue number2
DOIs
Publication statusPublished - Dec 1 2016

Fingerprint

Neural Crest
Glial Cell Line-Derived Neurotrophic Factor
Endothelin-3
Small Intestine
Large Intestine
Neurons
Hirschsprung Disease
Nerve Growth Factors
Therapeutics
Cell- and Tissue-Based Therapy
Intestines
Cell Movement
Culture Media
Colon
Stem Cells
Population

Keywords

  • Endothelin-3
  • Enteric nervous system
  • Enteric neural stem cells
  • Glial cell line-derived neurotrophic factor
  • Neurospheres

ASJC Scopus subject areas

  • Surgery

Cite this

Cheng, L. S., Graham, H. K., Pan, W. H., Nagy, N., Carreon-Rodriguez, A., Goldstein, A. M., & Hotta, R. (2016). Optimizing neurogenic potential of enteric neurospheres for treatment of neurointestinal diseases. Journal of Surgical Research, 206(2), 451-459. https://doi.org/10.1016/j.jss.2016.08.035

Optimizing neurogenic potential of enteric neurospheres for treatment of neurointestinal diseases. / Cheng, Lily S.; Graham, Hannah K.; Pan, Wei Hua; Nagy, N.; Carreon-Rodriguez, Alfonso; Goldstein, Allan M.; Hotta, Ryo.

In: Journal of Surgical Research, Vol. 206, No. 2, 01.12.2016, p. 451-459.

Research output: Contribution to journalArticle

Cheng, LS, Graham, HK, Pan, WH, Nagy, N, Carreon-Rodriguez, A, Goldstein, AM & Hotta, R 2016, 'Optimizing neurogenic potential of enteric neurospheres for treatment of neurointestinal diseases', Journal of Surgical Research, vol. 206, no. 2, pp. 451-459. https://doi.org/10.1016/j.jss.2016.08.035
Cheng, Lily S. ; Graham, Hannah K. ; Pan, Wei Hua ; Nagy, N. ; Carreon-Rodriguez, Alfonso ; Goldstein, Allan M. ; Hotta, Ryo. / Optimizing neurogenic potential of enteric neurospheres for treatment of neurointestinal diseases. In: Journal of Surgical Research. 2016 ; Vol. 206, No. 2. pp. 451-459.
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abstract = "Background Enteric neurospheres derived from postnatal intestine represent a promising avenue for cell replacement therapy to treat Hirschsprung disease and other neurointestinal diseases. We describe a simple method to improve the neuronal yield of spontaneously formed gut-derived neurospheres. Materials and methods Enteric neurospheres were formed from the small and large intestines of mouse and human subjects. Neurosphere size, neural crest cell content, cell migration, neuronal differentiation, and neuronal proliferation in culture were analyzed. The effect of supplemental neurotrophic factors, including glial cell line-derived neurotrophic factor (GDNF) and endothelin-3, was also assessed. Results Mouse small intestine–derived neurospheres contained significantly more P75-expressing neural crest-derived cells (49.9 ± 15.3{\%} versus 21.6 ± 11.9{\%}, P < 0.05) and gave rise to significantly more Tuj1-expressing neurons than colon-derived neurospheres (69.9 ± 8.6{\%} versus 46.2 ± 15.6{\%}, P < 0.05). A similar pattern was seen in neurospheres isolated from human small and large intestine (32.6 ± 17.5{\%} versus 10.2 ± 8.2{\%} neural crest cells, P < 0.05; 29.7 ± 16.4{\%} versus 16.0 ± 13.5{\%} enteric neurons, P < 0.05). The addition of GDNF to the culture media further improved the neurogenic potential of small intestinal neurospheres (75.9 ± 4.0{\%} versus 67.8 ± 5.8{\%}, P < 0.05) whereas endothelin-3 had no effect. Conclusions Enteric neurospheres formed from small intestine and supplemented with GDNF yield an enriched population of neural crest-derived progenitor cells and give rise to a high density of enteric neurons.",
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AU - Graham, Hannah K.

AU - Pan, Wei Hua

AU - Nagy, N.

AU - Carreon-Rodriguez, Alfonso

AU - Goldstein, Allan M.

AU - Hotta, Ryo

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N2 - Background Enteric neurospheres derived from postnatal intestine represent a promising avenue for cell replacement therapy to treat Hirschsprung disease and other neurointestinal diseases. We describe a simple method to improve the neuronal yield of spontaneously formed gut-derived neurospheres. Materials and methods Enteric neurospheres were formed from the small and large intestines of mouse and human subjects. Neurosphere size, neural crest cell content, cell migration, neuronal differentiation, and neuronal proliferation in culture were analyzed. The effect of supplemental neurotrophic factors, including glial cell line-derived neurotrophic factor (GDNF) and endothelin-3, was also assessed. Results Mouse small intestine–derived neurospheres contained significantly more P75-expressing neural crest-derived cells (49.9 ± 15.3% versus 21.6 ± 11.9%, P < 0.05) and gave rise to significantly more Tuj1-expressing neurons than colon-derived neurospheres (69.9 ± 8.6% versus 46.2 ± 15.6%, P < 0.05). A similar pattern was seen in neurospheres isolated from human small and large intestine (32.6 ± 17.5% versus 10.2 ± 8.2% neural crest cells, P < 0.05; 29.7 ± 16.4% versus 16.0 ± 13.5% enteric neurons, P < 0.05). The addition of GDNF to the culture media further improved the neurogenic potential of small intestinal neurospheres (75.9 ± 4.0% versus 67.8 ± 5.8%, P < 0.05) whereas endothelin-3 had no effect. Conclusions Enteric neurospheres formed from small intestine and supplemented with GDNF yield an enriched population of neural crest-derived progenitor cells and give rise to a high density of enteric neurons.

AB - Background Enteric neurospheres derived from postnatal intestine represent a promising avenue for cell replacement therapy to treat Hirschsprung disease and other neurointestinal diseases. We describe a simple method to improve the neuronal yield of spontaneously formed gut-derived neurospheres. Materials and methods Enteric neurospheres were formed from the small and large intestines of mouse and human subjects. Neurosphere size, neural crest cell content, cell migration, neuronal differentiation, and neuronal proliferation in culture were analyzed. The effect of supplemental neurotrophic factors, including glial cell line-derived neurotrophic factor (GDNF) and endothelin-3, was also assessed. Results Mouse small intestine–derived neurospheres contained significantly more P75-expressing neural crest-derived cells (49.9 ± 15.3% versus 21.6 ± 11.9%, P < 0.05) and gave rise to significantly more Tuj1-expressing neurons than colon-derived neurospheres (69.9 ± 8.6% versus 46.2 ± 15.6%, P < 0.05). A similar pattern was seen in neurospheres isolated from human small and large intestine (32.6 ± 17.5% versus 10.2 ± 8.2% neural crest cells, P < 0.05; 29.7 ± 16.4% versus 16.0 ± 13.5% enteric neurons, P < 0.05). The addition of GDNF to the culture media further improved the neurogenic potential of small intestinal neurospheres (75.9 ± 4.0% versus 67.8 ± 5.8%, P < 0.05) whereas endothelin-3 had no effect. Conclusions Enteric neurospheres formed from small intestine and supplemented with GDNF yield an enriched population of neural crest-derived progenitor cells and give rise to a high density of enteric neurons.

KW - Endothelin-3

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KW - Enteric neural stem cells

KW - Glial cell line-derived neurotrophic factor

KW - Neurospheres

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