Characterization of Fetal and Postnatal Enteric Neuronal Cell Lines With Improvement in Intestinal Neural Function

Anitha, Mallappa; Joseph, Irene; Ding, Xiaokun; Torre, Enrique; Sawchuk, Michael; Mwangi, Simon M.; Hochman, Shawn; Sitaraman, Shanthi V.; Anania, Frank A.; Srinivasan, Shanthi

doi:10.1053/j.gastro.2008.02.018

Cited by 68 publications

(78 citation statements)

References 25 publications

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“…p75-expressing cells were isolated from stomach and small and large bowel of E14.5 Sprague-Dawley rat embryos (10 -12 embryos) by magnetic bead immunoselection using a monoclonal antibody to p75NTR (10 g/ml) as previously described (18,28). The IM-FEN cell line was established and characterized as described previously (4). Following immunoselection, Trypan blue-excluding cells were plated onto a plastic 75-cm 2 cell culture flask in modified N2 medium (18) containing GDNF (100 ng/ml), 10% FBS, and 20 U/ml of recombinant mouse IFN-␥ and were cultured in a humidified tissue culture incubator containing 10% CO 2 at permissive temperature, 33°C.…”

Section: Methodsmentioning

confidence: 99%

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BMP2 promotes differentiation of nitrergic and catecholaminergic enteric neurons through a Smad1-dependent pathway

Anitha

Shahnavaz

Qayed

et al. 2010

American Journal of Physiology-Gastrointestinal and Liver Physi

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The bone morphogenetic protein (BMP) family is a class of transforming growth factor (TGF-␤) superfamily molecules that have been implicated in neuronal differentiation. We studied the effects of BMP2 and glial cell line-derived neurotrophic factor (GDNF) on inducing differentiation of enteric neurons and the signal transduction pathways involved. Studies were performed using a novel murine fetal enteric neuronal cell line (IM-FEN) and primary enteric neurons. Enteric neurons were cultured in the presence of vehicle, GDNF (100 ng/ml), BMP2 (10 ng/ml), or both (GDNF ϩ BMP2), and differentiation was assessed by neurite length, markers of neuronal differentiation (neurofilament medium polypeptide and ␤-III-tubulin), and neurotransmitter expression [neuropeptide Y (NPY), neuronal nitric oxide synthase (nNOS), tyrosine hydroxylase (TH), choline acetyltransferase (ChAT) and Substance P]. BMP2 increased the differentiation of enteric neurons compared with vehicle and GDNF-treated neurons (P Ͻ 0.001). BMP2 increased the expression of the mature neuronal markers (P Ͻ 0.05). BMP2 promoted differentiation of NPY-, nNOS-, and TH-expressing neurons (P Ͻ 0.001) but had no effect on the expression of cholinergic neurons (ChAT, Substance P). Neurons cultured in the presence of BMP2 have higher numbers of TH-expressing neurons after exposure to 1-methyl 4-phenylpyridinium (MPP ϩ ) compared with those cultured with MPP ϩ alone (P Ͻ 0.01). The Smad signal transduction pathway has been implicated in TGF-␤ signaling. BMP2 induced phosphorylation of Smad1, and the effects of BMP2 on differentiation of enteric neurons were significantly reduced in the presence of Smad1 siRNA, implicating the role of Smad1 in BMP2-induced differentiation. The effects of BMP2 on catecholaminergic neurons may have therapeutic implications in gastrointestinal motility disturbances.immorto; glial cell line-derived neurotrophic factor; bone morphogenic protein 2; Smad1 ENTERIC NEURONS ARE DERIVED from vagal, sacral, and upper thoracic neural crest cells that migrate into the gut, where they proliferate and then differentiate into a variety of distinct cell types (2,17,22,23). Several growth factors influence the proliferation, differentiation, and survival of the enteric nervous system (ENS), including glial cell line-derived neurotrophic factor (GDNF), neurturin, neurotrophin-3, and Sonic hedgehog (14, 18). Bone morphogenetic proteins (BMPs), members of the transforming growth factor-␤ (TGF-␤) superfamily, are regulatory molecules that play critical roles in the formation and regionalization of the gut before its colonization by crest-derived cells (26,27). BMP2 and 4 continue to be expressed in the fetal gut and regulate the specification, migration, differentiation, and aggregation of enteric neurons (12,13,15,24).We have recently developed an enteric neuronal cell line that has characteristics similar to primary enteric neurons (4). We sought to determine the types of growth factors that would result in differentiation of this novel enteric neuronal cel...

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Section: Methodsmentioning

confidence: 99%

“…We have recently developed an enteric neuronal cell line that has characteristics similar to primary enteric neurons (4). We sought to determine the types of growth factors that would result in differentiation of this novel enteric neuronal cell line.…”

mentioning

confidence: 99%

BMP2 promotes differentiation of nitrergic and catecholaminergic enteric neurons through a Smad1-dependent pathway

Anitha

Shahnavaz

Qayed

et al. 2010

American Journal of Physiology-Gastrointestinal and Liver Physi

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“…Furthermore, molecules produced by the gut mesenchyme are essential for the normal development of the ENS (12,(25)(26)(27), but it is unclear whether these factors are expressed at sufficient levels in the postnatal bowel to permit the development of enteric neurons from progenitors. Previous studies have transplanted CNS neural stem cells, ENS stem/progenitor cells, or ENS cell lines into the gut of postnatal animals in vivo (4,(28)(29)(30)(31)(32)(33) or grown cocultures between stem/progenitor cells and the muscle of postnatal human gut (13), but the extent of migration, and whether the graft-derived neurons have the electrophysiological properties of enteric neurons and are incorporated into the neuronal circuitry, have not been determined.…”

Section: Introductionmentioning

confidence: 99%

Transplanted progenitors generate functional enteric neurons in the postnatal colon

Hotta

Stamp²,

Foong³

et al. 2013

J. Clin. Invest.

149

207

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Cell therapy has the potential to treat gastrointestinal motility disorders caused by diseases of the enteric nervous system. Many studies have demonstrated that various stem/progenitor cells can give rise to functional neurons in the embryonic gut; however, it is not yet known whether transplanted neural progenitor cells can migrate, proliferate, and generate functional neurons in the postnatal bowel in vivo. We transplanted neurospheres generated from fetal and postnatal intestinal neural crest-derived cells into the colon of postnatal mice. The neurosphere-derived cells migrated, proliferated, and generated neurons and glial cells that formed ganglion-like clusters within the recipient colon. Graft-derived neurons exhibited morphological, neurochemical, and electrophysiological characteristics similar to those of enteric neurons; they received synaptic inputs; and their neurites projected to muscle layers and the enteric ganglia of the recipient mice. These findings show that transplanted enteric neural progenitor cells can generate functional enteric neurons in the postnatal bowel and advances the notion that cell therapy is a promising strategy for enteric neuropathies.

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“…The immortal postnatal enteric neuronal (IM-PEN) cell line 37 was seeded onto 6-well plates with modified N2 medium containing glial cell line-derived neurotrophic factor (100 ng/mL), 10% fetal bovine serum, and 20 U/mL of recombinant mouse interferon-g, and were cultured in a humidified tissue culture incubator containing 10% CO 2 at permissive temperature, 33 C. After 48 hours the medium was changed to Neurobasal-A medium containing B-27 serum-free supplement, 1 mmol/L glutamine, 1% fetal bovine serum, GDNF (100 ng/mL), and the plates were transferred to an atmosphere of 5% CO 2 at 39 C. Palmitate was used in 0.5-1 mmol/L concentrations, which is within the physiologically increased limits observed in human beings and animals, and can induce insulin resistance and hyperlipidemia-associated complications. 38 Palmitate was dissolved in isopropanol to obtain a stock concentration of 100 mmol/L.…”

Section: Immortal Postnatal Enteric Neuronal Cell Line Culturementioning

confidence: 99%

26 Intestinal Dysbiosis Contributes to the Delayed Gastrointestinal Transit in High Fat Diet Fed Mice

Anitha¹,

Nezami²,

Mwangi³

et al. 2014

Gastroenterology

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Characterization of Fetal and Postnatal Enteric Neuronal Cell Lines With Improvement in Intestinal Neural Function

Cited by 68 publications

References 25 publications

BMP2 promotes differentiation of nitrergic and catecholaminergic enteric neurons through a Smad1-dependent pathway

BMP2 promotes differentiation of nitrergic and catecholaminergic enteric neurons through a Smad1-dependent pathway

Transplanted progenitors generate functional enteric neurons in the postnatal colon

26 Intestinal Dysbiosis Contributes to the Delayed Gastrointestinal Transit in High Fat Diet Fed Mice

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