Journal
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
Volume 45, Issue 3-4, Pages 106-110Publisher
SPRINGER
DOI: 10.1007/s11626-009-9176-7
Keywords
Insect midgut; Primary cultures; Stem cells; Differentiated cells
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This protocol describes the preparation of primary cell cultures from Lepidopteran midgut. These cultures have been used to identify factors that control midgut growth and differentiation, cell responses to these factors, effects of toxins on midgut growth, and the regulation of cell physiology. The protocol is divided into (1) procedures for cell collection, (2) composition of the culture, and (3) assay methods used for cell health, proliferation, and differentiation. Collection and setup require 4-6 h. Once established, a culture can survive several months at 25A degrees C, be kept a year or longer at 4A degrees C, or be frozen for indefinite storage.
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