title: A continuous lineage of rat adenohypophysis stromal cells: characterisation and effects on GH(3)B(6) prolactin-secreting cell behaviour.
abstract: Although some studies have shown a possible modulation of the stroma on the hormonal secretion, it is not clear as to what are the requirements for these cellular interactions. In the present work, a homogeneous and continuous lineage of rat adenohypophysis stromal cells (APS9 cells) obtained from rat adenohypophysis primary culture was established. Using immunocytochemical methods and electron microscopy, we have characterised APS9 cells as elongated fibroblastoid-like cells with intercellular contacts, expressing alpha-smooth muscle actin, type IV collagen and laminin. By biochemical procedures, higher amounts of chondroitin sulphate and heparan sulphate were found in the pericellular and extracellular compartments of APS9 cell culture. In order to evaluate the possible effects of APS9 cell on GH(3)B(6) prolactin-secreting cell survival and/or proliferation, we established co-culture and proliferation assays. When GH(3)B(6) cells were cultivated on APS9 cell substrate, they displayed an organisation of many cellular cords strongly attached and covering all the stromal cell area, establishing punctual interactions or extensive surface associations between adjacent cells. Prolactin immunoreactivity appeared to be more scattered throughout the cytoplasm and accumulated in its periphery. When plated on glass coverslips, on newborn rat skin fibroblasts, on murine haematopoietic bone marrow stroma cell line or on murine foetal liver stroma cell line, GH(3)B(6) cells changed their organisation and presented a decrease in cell number and adherence to the substrate. Our results showed that the APS9 cell/GH(3)B(6) cell interactions favour cell growth and probably PRL secretion, and raises questions about the specificity of different organs and/or animal species stromas on the hormone secretion.
authors.affiliation: Departamento de Histologia e Embriologia, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Cidade Universitária - Ilha do Fundão 21941-590, Rio de Janeiro, Brazil.
authors.forename: Leandro M
authors.initials: LM
authors.lastname: Alves
authors.forename: Cláudio C
authors.initials: CC
authors.lastname: Werneck
authors.forename: Rita de Cássia L
authors.initials: Rde C
authors.lastname: Martins Rd
authors.forename: Luiz Cláudio F
authors.initials: LC
authors.lastname: Silva
authors.forename: Marília
authors.initials: M
authors.lastname: Taffarel
authors.forename: Radovan
authors.initials: R
authors.lastname: Borojevic
authors.forename: Luiz E
authors.initials: LE
authors.lastname: Nasciutti
mesh_terms: D000818:Animals; D002450:Cell Communication / Q000502:physiology*; D002455:Cell Division; D002460:Cell Line; D019070:Cell Lineage; D048429:Cell Size; D018920:Coculture Techniques; D008297:Male; D051379:Mice; D008856:Microscopy, Fluorescence; D010903:Pituitary Gland, Anterior / Q000166:cytology* / Q000502:physiology; D011388:Prolactin / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D017154:Stromal Cells / Q000166:cytology* / Q000502:physiology
chemical_list: D011388:Prolactin