title: Three-dimensional Alginate-bead Culture of Human Pituitary Adenoma Cells.
abstract: A three-dimensional culture method is described in which primary pituitary adenoma cells are grown in alginate beads. Alginate is a polymer derived from brown sea algae. Briefly, the tumor tissue is cut into small pieces and submitted to an enzymatic digestion with collagenase and trypsin. Next, a cell suspension is obtained. The tumor cell suspension is mixed with 1.2% sodium alginate and dropped into a CaCl2 solution, and the alginate/cell suspension is gelled on contact with the CaCl2 to form spherical beads. The cells embedded in the alginate beads are supplied with nutrients provided by the culture media enriched with 20% FBS. Three-dimensional culture in alginate beads maintains the viability of adenoma cells for long periods of time, up to four months. Moreover, the cells can be liberated from the alginate by washing the beads with sodium citrate and seeded on glass coverslips for further immunocytochemical analyses. The use of a cell culture model allows for the fixation and visualization of the actin cytoskeleton with minimal disorganization. In summary, alginate beads provide a reliable culture system for the maintenance of pituitary adenoma cells.
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, Center for Research and Advanced Studies (CINVESTAV-IPN).
authors.forename: Dulce
authors.initials: D
authors.lastname: Avila-Rodríguez
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, Center for Research and Advanced Studies (CINVESTAV-IPN).
authors.forename: Karina
authors.initials: K
authors.lastname: Paisano-Cerón
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, Center for Research and Advanced Studies (CINVESTAV-IPN).
authors.forename: Irene
authors.initials: I
authors.lastname: Valdovinos-Ramírez
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, Center for Research and Advanced Studies (CINVESTAV-IPN).
authors.forename: Carmen
authors.initials: C
authors.lastname: Solano-Agama
authors.affiliation: Department of Experimental Pathology, Nacional Institute of Neurology and Neurosurgery.
authors.forename: Alma
authors.initials: A
authors.lastname: Ortiz-Plata
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, Center for Research and Advanced Studies (CINVESTAV-IPN); mmendoza@fisio.cinvestav.mx.
authors.forename: María E
authors.initials: ME
authors.lastname: Mendoza-Garrido
mesh_terms: D008841:Actin Cytoskeleton / Q000473:pathology; D000464:Alginates; D002122:Calcium Chloride / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods*; D060326:Chemistry Techniques, Synthetic / Q000379:methods; D003470:Culture Media; D004798:Enzymes; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D008863:Microspheres; D010911:Pituitary Neoplasms / Q000473:pathology*
chemical_list: D000464:Alginates; D003470:Culture Media; D004798:Enzymes; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D002122:Calcium Chloride