title: Two methods for establishing primary human endometrial stromal cells from hysterectomy specimens.
abstract: Many efforts have been devoted to establish in vitro cell culture systems. These systems are designed to model a vast number of in vivo processes. Cell culture systems arising from human endometrial samples are no exception. Applications range from normal cyclic physiological processes to endometrial pathologies such as gynecological cancers, infectious diseases, and reproductive deficiencies. Here, we provide two methods for establishing primary endometrial stromal cells from surgically resected endometrial hysterectomy specimens. The first method is referred to as "the scraping method" and incorporates mechanical scraping using surgical or razor blades whereas the second method is termed "the trypsin method." This latter method uses the enzymatic activity of trypsin to promote the separation of cells and primary cell outgrowth. We illustrate step-by-step methodology through digital images and microscopy. We also provide examples for validating endometrial stromal cell lines via quantitative real time polymerase chain reactions (qPCR) and immunofluorescence (IF).
authors.affiliation: Department of Pathology, University of Virginia.
authors.forename: Kasey
authors.initials: K
authors.lastname: Jividen
authors.affiliation: Department of Pathology, University of Virginia.
authors.forename: Mercedeh Javanbakht
authors.initials: MJ
authors.lastname: Movassagh
authors.affiliation: Department of Obstetrics & Gynecology, University of Virginia.
authors.forename: Amir
authors.initials: A
authors.lastname: Jazaeri
authors.affiliation: Department of Pathology, University of Virginia; hl9r@virginia.edu.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D004717:Endometrium / Q000166:cytology* / Q000601:surgery; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007044:Hysterectomy; D060888:Real-Time Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology*