Magnetic cell sorting for parietal cell purification using a new monoclonal antibody without influence on cell function.
Conventional isolation protocols for the parietal type of the gastric mucosal cell population are based on physical criteria such as cell size and cell density. Time-consuming centrifugation techniques are required, which decrease the overall yield and exert a negative influence on the functional integrity of purified cells. As an alternative, we now have developed a phenotype-dependent magnetic selection method. A monoclonal antibody (P1.4D5) was raised against rat parietal cells. After immunohistochemical, biochemical and functional characterization, P1.4D5 was used for rapid antibody-mediated magnetic cell sorting (MACS) of enzymatically dispersed gastric mucosal cells, which resulted in a 2.2- to 3-fold enrichment of parietal cells. Characterization of P1.4D5 demonstrated its applicability for parietal cell-specific immunostaining of formalin treated and paraffin-embedded tissue samples and cryosections from both rat and man. In vitro studies of the 14C-aminopyrine accumulation showed no intrinsic effect of P1.4D5 on the acid production of the parietal cell or on the histamine-stimulated acid production. This absence of negative influence on the parietal cell function qualifies P1.4D5 for immunomagnetic cell sorting procedures.
Department of Internal Medicine I, Eberhard-Karls-University, Tübingen, Germany. florian.graepler@unituebingen.de
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Graepler
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Lauer
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Gregor
D000632:Aminopyrine / Q000493:pharmacokinetics; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry; D006632:Histamine / Q000494:pharmacology; D006651:Histocytochemistry; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008280:Magnetics; D010295:Parietal Cells, Gastric / Q000166:cytology*; D051381:Rats
D000911:Antibodies, Monoclonal; D000632:Aminopyrine; D006632:Histamine