The enteric neural crest progressively loses capacity to form enteric nervous system

• Published in: Developmental biology Vol. 446; no. 1; pp. 34 - 42
• Main Authors: Zhang, Dongcheng, Rollo, Benjamin N., Nagy, Nandor, Stamp, Lincon, Newgreen, Donald F.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.02.2019
• Subjects: Animals; Cell Differentiation; Cell Movement; Cells, Cultured; Chick Embryo; Chickens; Chorioallantoic Membrane - transplantation; Coturnix; Digestive System - cytology; Digestive System - embryology; Digestive System - metabolism; Enteric Nervous System - cytology; Enteric Nervous System - embryology; Enteric Nervous System - metabolism; Intestine, Small - cytology; Intestine, Small - embryology; Intestine, Small - innervation; Neural Crest - cytology; Neural Crest - embryology; Neural Crest - metabolism; Neuroglia - cytology; Neuroglia - metabolism; Neurons - cytology; Neurons - metabolism; Organ Culture Techniques
• ISSN: 0012-1606; 1095-564X
• DOI: 10.1016/j.ydbio.2018.11.017

Cells of the vagal neural crest (NC) form most of the enteric nervous system (ENS) by a colonising wave in the embryonic gut, with high cell proliferation and differentiation. Enteric neuropathies have an ENS deficit and cell replacement has been suggested as therapy. This would be performed post-natally, which raises the question of whether the ENS cell population retains its initial ENS-forming potential with age. We tested this on the avian model in organ culture in vitro (3 days) using recipient aneural chick midgut/hindgut combined with ENS-donor quail midgut or hindgut of ages QE5 to QE10. ENS cells from young donor tissues (≤ QE6) avidly colonised the aneural recipient, but this capacity dropped rapidly 2–3 days after the transit of the ENS cell wavefront. This loss in capability was autonomous to the ENS population since a similar decline was observed in ENS cells isolated by HNK1 FACS. Using QE5, 6, 8 and 10 midgut donors and extending the time of assay to 8 days in chorio-allantoic membrane grafts did not produce ‘catch up’ colonisation. NC-derived cells were counted in dissociated quail embryo gut and in transverse sections of chick embryo gut using NC, neuron and glial marker antibodies. This showed that the decline in ENS-forming ability correlated with a decrease in proportion of ENS cells lacking both neuronal and glial differentiation markers, but there were still large numbers of such cells even at stages with low colonisation ability. Moreover, ENS cells in small numbers from young donors were far superior in colonisation ability to larger numbers of apparently undifferentiated cells from older donors. This suggests that the decline of ENS-forming ability has both quantitative and qualitative aspects. In this case, ENS cells for cell therapies should aim to replicate the embryonic ENS stage rather than using post-natal ENS stem/progenitor cells. •Avian wavefront embryonic enteric neural crest cells efficiently form ENS.•This efficiency persists only briefly, for about 2 days, then drops dramatically.•The drop in ENS-forming efficiency is innate to the enteric neural crest cells.•This parallels a decrease in proportion of undifferentiated ENS cells.•But this occurs despite an increase in the number of undifferentiated ENS cells.