Abstract
CXCR4, the stromal cell-derived factor-1 receptor, plays an important role in the migration of hematopoietic progenitor/stem cells to injured and inflamed areas. Noninvasive cell tracking methods could be useful for monitoring cell fate. Therefore, in this study, we evaluated the efficacy of an intravenous infusion of genetically engineered mesenchymal stem cells (MSCs) overexpressing CXC chemokine receptor 4 (CXCR4) to home to the tumor, by optical imaging. We constructed a retroviral vector containing CXCR with dual reporter genes, eGFP and Fluc2, under the control of an EF1α promoter (pBABE-EF1α-CXCR4-eGFP-IRES-Fluc2). We also developed an eGFP-Fluc2 construct in the Retro-X retroviral vector (Retro-X-eGFP-Fluc2). MSCs were transduced with retroviruses to generate CXCR4-overexpressing MSCs (MSC-CXCR4/Fluc2) and MSCs (MSC/Fluc2). CXCR4 mRNA and protein expression was confirmed by RT-PCR and Western blotting, respectively, and it was higher in MSC-CXCR4/Fluc2 than in naive MSCs. eGFP expression was confirmed by confocal microscopy. The transfected MSC-CXCR4/Fluc2 cells showed higher migratory capacity than naive MSCs observed in Transwell migration assay. The in vivo migration of CXCR4-overexpressing MSCs to MDAMB231/Rluc tumor model by BLI imaging was also confirmed. Intravenous delivery of genetically modified MSCs overexpressing CXCR4 with a Fluc2 reporter gene may be a useful, noninvasive BLI imaging tool for tracking cell fate.
Highlights
IntroductionThere are 1.38 million women with breast cancer [1]. Breast cancer is one of the most common cancers among women, especially in less developed countries and the second leading cause of cancer death [2]
Worldwide, there are 1.38 million women with breast cancer [1]
Transduced mesenchymal stem cells (MSCs)-CXC chemokine receptor 4 (CXCR4)/Fluc2 cells were generated by retroviral transfection, and eGFP-positive cells were sorted by FACS Aria III
Summary
There are 1.38 million women with breast cancer [1]. Breast cancer is one of the most common cancers among women, especially in less developed countries and the second leading cause of cancer death [2]. Triple negative breast cancer (TNBC) is a subtype of tumors that do not clinically express human epidermal growth factor receptor 2 (HER-2), progesterone receptor (PR), or estrogen receptor (ER). Mesenchymal stem cell- (MSC-) based homing studies may lead to a better approach for treating such breast cancers. MSCs are most commonly isolated from bone marrow [12], they are isolated from other tissues, including adipose tissue [13, 14], Stem Cells International the placenta [15], amniotic fluid [16], and umbilical cord blood [17, 18] Owing to their accessibility and convenient expansion protocols, MSCs have been recognized as promising candidates for cellular therapy. In the current study, we established CXCR4-overexpressing MSCs, containing a Fluc reporter gene, and evaluated their in vivo migrating efficiency to triple negative breast cancer by BLI
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