title: Primary culture of porcine mammary epithelial cells as a model system for evaluation of milk protein expression.
abstract: Porcine mammary epithelial cells were isolated to culture on collagen gel followed by gel floating treatment to evaluate differentiation under the culture conditions of serum-free medium, supplemented with combinations of insulin, hydrocortisone, and prolactin. After the culture period, the mammary cells attached to the collagen gels were recovered to observe expression of beta-casein, beta-lactoglobulin, and lactoferrin by reverse transcriptase polymeric chain reaction method. Expression of beta-casein was observed in the presence of insulin, hydrocortisone, and prolactin whereas transcription of beta-lactoglobulin and lactoferrin occured irrespective of hydrocortisone and prolactin. Immunoblot analysis demonstrated synthesis and secretion of lactoferrin in the fraction of recovered cells and the culture medium.
authors.affiliation: Division of Bioresources and Product Science, The Graduate School of Agriculture, Hokkaido University, Sapporo, Japan. kumura@anim.agr.hokudai.ac.jp
authors.forename: H
authors.initials: H
authors.lastname: Kumura
authors.forename: A
authors.initials: A
authors.lastname: Tanaka
authors.forename: Y
authors.initials: Y
authors.lastname: Abo
authors.forename: S
authors.initials: S
authors.lastname: Yui
authors.forename: K
authors.initials: K
authors.lastname: Shimazaki
authors.forename: E
authors.initials: E
authors.lastname: Kobayashi
authors.forename: K
authors.initials: K
authors.lastname: Sayama
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology; D005260:Female; D006854:Hydrocortisone / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D008321:Mammary Glands, Animal / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008894:Milk Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011388:Prolactin / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine
chemical_list: D016895:Culture Media, Serum-Free; D007328:Insulin; D008894:Milk Proteins; D011388:Prolactin; D006854:Hydrocortisone