Isogenic enteric neural progenitor cells can replace missing neurons and glia in mice with Hirschsprung disease.

BACKGROUND: Transplanting autologous patient-derived enteric neuronal stem/progenitor cells (ENSCs) is an innovative approach to replacing missing enteric neurons in patients with Hirschsprung disease (HSCR). Using autologous cells eliminates immunologic and ethical concerns raised by other cell sources. However, whether postnatal aganglionic bowel is permissive for transplanted ENSCs and whether ENSCs from HSCR patients can be successfully isolated, cultured, and transplanted in vivo remains unknown. METHODS: ENSCs isolated from the ganglionic intestine of Ednrb(-/-) mice (HSCR-ENSCs) were characterized immunohistochemically and evaluated for their capacity to proliferate and differentiate in vitro. Fluorescently labeled ENSCs were co-cultured ex vivo with aganglionic Ednrb(-/-) colon. For in vivo transplantation, HSCR-ENSCs were labeled with lentivirus expressing green fluorescent protein (GFP) and implanted into aganglionic embryonic chick gut in ovo and postnatal aganglionic Ednrb(-/-) rectum in vivo. KEY RESULTS: HSCR-ENSCs maintain normal capacity self-renewal and neuronal differentiation. Moreover, the Ednrb(-/-) aganglionic environment is permissive to engraftment by wild-type ENSCs ex vivo and supports migratrion and neuroglial differentiation of these cells following transplantation in vivo. Lentiviral GFP-labeled HSCR-ENSCs populated embryonic chick hindgut and postnatal colon of Ednrb(-/-) HSCR, with cells populating the intermuscular layer and forming enteric neurons and glia. CONCLUSIONS & INFERENCES: ENSCs can be isolated and cultured from mice with HSCR, and transplanted into the aganglionic bowel of HSCR littermates to generate enteric neuronal networks. These results in an isogenic model establish the potential of using autologous-derived stem cells to treat HSCR and other intestinal neuropathies.

Endoscopic delivery of enteric neural stem cells to treat Hirschsprung disease.

BACKGROUND: Transplantation of enteric neural stem cells (ENSC) holds promise as a potential therapy for enteric neuropathies, including Hirschsprung disease. Delivery of transplantable cells via laparotomy has been described, but we propose a novel, minimally invasive endoscopic method of cell delivery. METHODS: Enteric neural stem cells for transplantation were cultured from dissociated gut of postnatal donor mice. Twelve recipient mice, including Ednrb(-/-) mice with distal colonic aganglionosis, underwent colonoscopic injection of ENSC under direct vision using a 30-gauge Hamilton needle passed through a rigid cystoureteroscope. Cell engraftment, survival, and neuroglial differentiation were studied 1-4 weeks after the procedure. KEY RESULTS: All recipient mice tolerated the procedure without complications and survived to sacrifice. Transplanted cells were found within the colonic wall in 9 of 12 recipient mice with differentiation into enteric neurons and glia. CONCLUSIONS & INFERENCES: Endoscopic injection of ENSC is a safe and reliable method for cell delivery, and can be used to deliver a large number of cells to a specific area of disease. This minimally invasive endoscopic approach may prove beneficial to future human applications of cell therapy for neurointestinal disease.

Nestin-expressing cells in the gut give rise to enteric neurons and glial cells.

BACKGROUND: Neuronal stem cells (NSCs) are promising for neurointestinal disease therapy. Although NSCs have been isolated from intestinal musclularis, their presence in mucosa has not been well described. Mucosa-derived NSCs are accessible endoscopically and could be used autologously. Brain-derived Nestin-positive NSCs are important in endogenous repair and plasticity. The aim was to isolate and characterize mucosa-derived NSCs, determine their relationship to Nestin-expressing cells and to demonstrate their capacity to produce neuroglial networks in vitro and in vivo. METHODS: Neurospheres were generated from periventricular brain, colonic muscularis (Musc), and mucosa-submucosa (MSM) of mice expressing green fluorescent protein (GFP) controlled by the Nestin promoter (Nestin-GFP). Neuronal stem cells were also grown as adherent colonies from intestinal mucosal organoids. Their differentiation potential was assessed using immunohistochemistry using glial and neuronal markers. Brain and gut-derived neurospheres were transplanted into explants of chick embryonic aneural hindgut to determine their fate. KEY RESULTS: Musc- and MSM-derived neurospheres expressed Nestin and gave rise to cells of neuronal, glial, and mesenchymal lineage. Although Nestin expression in tissue was mostly limited to glia co-labelled with glial fibrillary acid protein (GFAP), neurosphere-derived neurons and glia both expressed Nestin in vitro, suggesting that Nestin+/GFAP+ glial cells may give rise to new neurons. Moreover, following transplantation into aneural colon, brain- and gut-derived NSCs were able to differentiate into neurons. CONCLUSIONS & INFERENCES: Nestin-expressing intestinal NSCs cells give rise to neurospheres, differentiate into neuronal, glial, and mesenchymal lineages in vitro, generate neurons in vivo and can be isolated from mucosa. Further studies are needed for exploring their potential for treating neuropathies.

Fatty acids and neurodevelopment.

Knowledge of the importance of docosahexaenoic acid (DHA), arachidonic acid (AA), and long-chain polyunsaturated fatty acids (LCPUFAs) in neurodevelopment was originally obtained from animal studies. These fatty acids are rapidly accreted in brain during the first postnatal year in animal and human infants, and they are found in high concentrations in breast milk. Reports of enhanced intellectual development in breast-fed children, and reports linking LCPUFA deficiency with neurodevelopmental disorders have stressed the physiological importance of DHA in visual and neural systems. In addition to high concentrations of fatty acids in breast milk, they are also present in fish and algae oil and have recently been added to infant formulas. Esterified poplyunsaturated fatty acids act in cellular membranes, in signal transduction, in neurotransmission, and in the formation of lipid rafts. Nonesterified polyunsaturated fatty acids can modulate gene expression and ion channel activities, thus becoming neuroprotective agents. The conversion of linoleic acid and alpha-linolenic acid into ARA and DHA have led to randomized clinical trials that have studied whether infant formulas supplemented with DHA or both DHA and ARA would enhance visual and cognitive development. This review gives an overview of fatty acids and neurodevelopment, focusing on the findings from these studies.

Shwachman-Diamond syndrome in a Mexican family.

Shwachman-Diamond Syndrome (SDS) is an inherited condition with multisystemic abnormalities including pancreatic exocrine dysfunction, neutropenia, short stature, and skeletal abnormalities. In this report, we describe the case of a 14-year-old female with a history of neutropenia, pancreatic exocrine insufficiency and pancreatic endocrine sufficiency, pancreatic lipomatosis (10), and the development of myeloid leukemia. Postmortem examination revealed a high probability of SDS. We also describe the clinical findings in the patient's six siblings, suggesting this as a familial form of SDS. Because the gene(s) responsible for this syndrome have not yet been identified, genetic confirmation is not yet possible. This is the first report in the literature of a Mexican family with probable SDS.

[Incidence of Helicobacter pylori infection in a cohort of infants in the State of Morelos]. / Incidencia de infección por Helicobacter pylori en una cohorte de lactantes en el estado de Morelos.

OBJECTIVE: To study the incidence of Helicobacter pylori infection in infants from the State of Morelos, Mexico. MATERIAL AND METHODS: A cohort of 110 healthy infants was studied between 1997 and 1999. Serum samples were collected from mothers and their infants at 2, 6, 18, and 24 months of life. All serum samples were tested for antibodies against Helicobacter pylori with the ELISA test. A questionnaire was used to collect socio-economic and clinical data. Associations among selected variables and Helicobacter pylori infection were determined using Fisher's exact test. RESULTS: Two thirds of mothers and six (5.5%) infants tested positive. Two of the six positive infants were born to positive mothers; both of them became negative before age two. The other four infants remained positive. Although not statistically significant, a vaginal birth and more than five people living in the household are possible risk factors for infant H. pylori infection. CONCLUSIONS: Despite the previously reported high prevalence of infection by H. pylori in Mexican children, in this population we found a low incidence of infection in infants up to two years of age. The English version of this paper is available at: http://www.insp.mx/salud/index.html.