Non-invasive exploration of neonatal gastric epithelium by using exfoliated epithelial cells.
OBJECTIVE
In preterm infants, exfoliated gastric epithelial cells can be retrieved from aspirates sampled through the naso-gastric feeding tube. Our aims were to determine (1) whether the recovery of exfoliated cells is feasible at any time from birth through the removal of the nasogastric tube, (2) whether they can be grown in culture in vitro, and (3) whether the physiological state of exfoliated cells expressing H+/K+ -ATPases reflects that of their counterparts remaining in situ at the surface of the gastric epithelium in neonatal rat pups.


METHODS
In infants, gastric fluid aspirates were collected weekly after birth or every 3 hours over 24-h periods, and related to clinical parameters (Biocollection PROG/09/18). In rat pups submitted to a single fasting/refeeding cycle, we explored circadian exfoliation with the cellular counter-parts in the gland. All samples were analyzed by confocal imaging and Enzyme-Linked Immunosorbent Assay.


RESULTS
Epithelial cells were identified by microscopy using membrane-bound anti-H+/K+ ATPases antibody, assessed for nucleus integrity, and the expression of selected proteins (autophagy, circadian clock). On 34 infants, the H+/K+-ATPase-positive cells were consistently found quiescent, regardless of gestational age and feeding schedule from day-5 of life to the day of removal of the naso-gastric tube. By logistic regression analysis, we did find a positive correlation between the intensity of exfoliation (cellular loss per sample) and the postnatal age (p<0.001). The H+/K+ ATPase-positive cells established in culture retained the expression of a biomarker of progenitor status (Pouf5F1-Oct4). In rat pups, the expression pattern of Survivin in H+/K+ ATPase-positive exfoliated cells paralleled that observed in cells remaining at the surface of the gastric gland.


CONCLUSIONS
Tracking parietal cells can improve clinical monitoring and understanding of the autophagic death via the phosphatidylinositol 3-kinase/Akt/survivin pathway.
Unité Mixte de Recherche-1280, Physiologie des Adaptations Nutritionnelles, Institut National Recherche Agronomique, Nantes, France. Bertrand.Kaeffer@univ-nantes.fr
Bertrand
B
Kaeffer
Arnaud
A
Legrand
Thomas
T
Moyon
Anne
A
Frondas-Chauty
Hélène
H
Billard
Omar
O
Guzman-Quevedo
Dominique
D
Darmaun
Jean-Christophe
JC
Rozé
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D004750:Enteral Nutrition; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D004848:Epithelium / Q000378:metabolism; D005260:Female; D005753:Gastric Mucosa / Q000166:cytology / Q000378:metabolism / Q000473:pathology; D005786:Gene Expression Regulation; D017506:H(+)-K(+)-Exchanging ATPase / Q000378:metabolism; D006760:Hospitalization; D006801:Humans; D007231:Infant, Newborn; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D011247:Pregnancy; D047928:Premature Birth / Q000378:metabolism / Q000473:pathology; D051381:Rats; D013234:Stem Cells / Q000166:cytology; D013270:Stomach / Q000166:cytology*; D013396:Suction; D000077022:Survivin; D013997:Time Factors
D015415:Biomarkers; C516989:Birc5 protein, rat; D008869:Microtubule-Associated Proteins; D000077022:Survivin; D017506:H(+)-K(+)-Exchanging ATPase