Separation of Gastric Mucosal Cells of Rat with Proteolytic Enzymes, Pronase and Trypsin, with Special Reference to the Collection, Morphology and Viability of the Generative Cells

Abstract

Methods to separate and collect gastric mucosal cells of the rat using proteolytic enzymes were devised. Pronase (1.0%) achieved better results than did trypsin (2.0%) in collecting single isolated cells with higher cell yields and viability. The cells dissociated with trypsin retained glandular structures as in situ. The measurement of radioactivity revealed that the incorporation of ³H-thymidine into generative cells was highest in the cell suspension collected by the second 15 min dissociation. It was concluded that the most effective method to obtain dissociated cells from the generative zone of the mucosa is to collect the cells dissociated with 1.0% pronase continuously for a period from 15 to 45 min after the start of dissociation. On autoradiographic analysis with ³H-thymidine, the ratio of generative cells was 10%, approximately 3 × 10⁶ cells, in the specimens.