title: Primary culture of insect midgut cells.
abstract: 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 25 degrees C, be kept a year or longer at 4 degrees C, or be frozen for indefinite storage.
authors.affiliation: Department of Anatomy, College of Medicine, Howard University, 520 W. St NW, Washington, DC 20059, USA. rhakim@mac.com
authors.forename: Raziel S
authors.initials: RS
authors.lastname: Hakim
authors.forename: Silvia
authors.initials: S
authors.lastname: Caccia
authors.forename: Marcia
authors.initials: M
authors.lastname: Loeb
authors.forename: Guy
authors.initials: G
authors.lastname: Smagghe
mesh_terms: D000818:Animals; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation; D002470:Cell Survival; D002478:Cells, Cultured; D004064:Digestive System / Q000166:cytology*; D007313:Insecta / Q000166:cytology*; D007814:Larva / Q000166:cytology; D008858:Microscopy, Phase-Contrast; D013234:Stem Cells / Q000166:cytology