Intraductal Injection of Lentivirus Vectors for Stably Introducing Genes into Rat Mammary Epithelial Cells in Vivo
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Intraductal Injection of Lentivirus Vectors for Stably Introducing Genes into Rat Mammary Epithelial Cells in Vivo Wen Bu 1,2 & Yi Li 1,2,3 Received: 29 July 2020 / Revised: 18 October 2020 / Accepted: 4 November 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Various retroviral and lentiviral vectors have been used for up-the-teat intraductal injection to deliver markers, oncogenes, and other genes into mammary epithelial cells in mice. These methods along with the large number of genetically engineered mouse lines have greatly helped us learn normal breast development and tumorigenesis. Rats are also valuable models for studying human breast development and cancer. However, genetically engineered rats are still uncommon, and previous reports of intraductal injection of retroviral vectors into rats appear to be inefficient in generating mammary tumors. Here, we report, and describe the method for, stably introducing marker genes and oncogenes into mammary glands in rats using intraductal injection of commonly used lentiviral vectors. This method can infect mammary epithelial cells efficiently, and the infected cells can initiate tumorigenesis, including estrogen receptor-positive and hormone-dependent tumors, which are the most common subtype of human breast cancer but are yet still difficult to model in mice. This technique provides another tool for studying formation, prevention, and treatment of breast cancer, especially estrogen receptor-positive breast cancer.
Introduction There are many mouse models of breast cancer, including conventional xenograft models, carcinogen-induced tumors, intraductal injection of breast tumor cells, genetically engineered mice, and genetic alterations of mammary epithelial cells by intraductal injection of viral vectors [1–8]. These models have greatly advanced our understanding of breast cancer formation, progression, and therapy and therapeutic resistance. However, a mouse mammary gland is quite dissimilar from a human breast: the mammary ductal tree in a sexually mature non-lactating mouse terminates in blunt ductules, and lobuloalveolar development towards terminal ductal lobular units (TDLUs) occurs only during pregnancy and largely regresses following involution. In contrast, the mammary ductal tree in a sexually mature non-lactating woman already ends in TDLUs that harbor extensive amounts of acini
* Yi Li [email protected] 1
Lester & Sue Smith Breast Center, Baylor College of Medicine, Houston, TX 77030, USA
2
Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030, USA
3
Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX 77030, USA
[9]. Furthermore, the mouse mammary stroma is largely comprised of adipose tissue with scanty connective tissue, while the human breast stroma contains dense amounts of collagenous connective tissue beside the adipose tissue [9, 10]. Compared to mammary glands of mice, those of rats are more similar to the human breast – in sexual
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