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Mouse Mammary Intraductal (MIND) Method for Transplantation of Patient Derived Primary DCIS Cells and Cell Lines.
Kittrell, Frances; Valdez, Kelli; Elsarraj, Hanan; Hong, Yan; Medina, Daniel; Behbod, Fariba.
Afiliação
  • Kittrell F; Department of Cell Biology, Baylor College of Medicine, University of Kansas Medical School, Kansas City, USA.
  • Valdez K; Department of Pathology and Laboratory Medicine, University of Kansas Medical School, Kansas City, USA.
  • Elsarraj H; Department of Pathology and Laboratory Medicine, University of Kansas Medical School, Kansas City, USA.
  • Hong Y; Department of Pathology and Laboratory Medicine, University of Kansas Medical School, Kansas City, USA.
  • Medina D; Department of Cell Biology, Baylor College of Medicine, University of Kansas Medical School, Kansas City, USA.
  • Behbod F; Department of Pathology and Laboratory Medicine, University of Kansas Medical School, Kansas City, USA.
Bio Protoc ; 6(5)2016 Mar 05.
Article em En | MEDLINE | ID: mdl-27446983
ABSTRACT
The MIND method involves intraductal injection of patient derived ductal carcinoma in situ (DCIS) cells and DCIS cell lines (MCF10DCIS.COM and SUM225) inside the mouse mammary ducts [Video 1 and Figure 1 in Behbod et al. (2009)]. This method mimics the normal environment of DCIS and facilitates study of the natural progression of human DCIS, i.e., their initial growth as carcinoma in situ within the ducts, followed by invasion into the stroma through the myoepithelial cell layer and basement membrane (Behbod et al., 2009; Valdez et al., 2011). In order to demonstrate that transplantation procedure is successful, the transplanted mammary glands may be excised as early as two weeks following intraductal injection of cells followed by Hematoxylin and Eosin (H&E) staining and/or immunofluorescence staining using human specific cytokeratin 5 and/or 19 [please see Figures 2-4 in Behbod et al. (2009)]. Additionally, the presence of trypan blue inside the mouse mammary ducts immediately following intraductal injection is the best indicator that the injection was successful (Video 1 starting at 433 sec).

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Bio Protoc Ano de publicação: 2016 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Bio Protoc Ano de publicação: 2016 Tipo de documento: Article País de afiliação: Estados Unidos