Studies Using Lifeline® Human Primary Cells for Transfection
Lifeline® offers a large number of primary cells for your research needs, many of which can be efficiently transfected. Importantly, Lifeline® cells can be used as normal control cells for various assays and are amenable to multiple transfection reagents. Our cells are particularly good models of normal cells compared with immortalized cell lines, as they lack the genetic alterations that are required for a cell to override cell cycle signals and become immortalized.
Yoshida et al. investigated the role of Thrombospondin-2 (TSP-2) in initimal hyperplasia, a condition that can develop following prosthetic arterial grafting. They had previously identified a significant upregulation of TSP-2 in initimal hyperplasia and hypothesized that it might contribute to increased cell proliferation and migration. To definitively investigate the physiological role of TSP-2 in aortic cells, the authors knocked down TSP-2 in Lifeline® Normal Human Aortic Smooth Muscle Cells (HAoSMCs) grown in VascuLife® basal medium with VascuLife® SMC LifeFactors. Unexpectedly, they found that TSP-2 did not alter cell proliferation or migration, but instead improved cell attachment. Their results suggest that regulating cell attachment might be an important role for TSP-2.
The authors report efficient knockdown of TSP-2 in Lifeline® HAoSMCs using HiPerFect transfection reagent (Qiagen Inc). In particular, TSP-2 protein was reduced to 10% of the wild-type levels and the knockdown was maintained in HAoSMCs for up to 30 days, indicating that these cells can readily tolerate transfection and siRNA-mediated protein knockdown.
Lewis et al. studied the effects of STAT3 antisense oligonucleotides on prostate and pancreatic cancer cell apoptosis. STAT3 is a transcription factor that is disrupted in various cancer types, where it mediates pro-tumorigenic pathways. The authors used antisense oligonucleotides that were complimentary to the genomic STAT3 consensus binding sequence, thereby blocking the ability of STAT3 to bind to its transcriptional targets. They found that STAT3 antisense oligonucleotides induced apoptosis in both prostate and pancreatic cancer cells, suggesting that STAT3 activity is important for tumor cell survival. In particular, they demonstrated that the 3’ end of the STAT3 consensus site was essential for induction of apoptosis in these cancer cells. Their results suggest that targeting STAT3 function in cancer might be a viable therapeutic opportunity.
The authors used Lifeline® primary Human Dermal Fibroblasts (HDFs) as a physiological normal control for their studies. They transfected Lifeline® HDFs with STAT3 antisense oligonucleotides using LipofectAMINE 2000 (Thermo Fisher Scientific) transfection reagent and found that the Lifeline® cells did not undergo apoptosis. These results demonstrate that the STAT3 antisense oligonucleotides are especially specific for the cancer cells tested and spare normal cells that might be neighboring a tumor.
Lifeline® Human Primary Cells
Lifeline® cell types that have been used successfully for transfection and gene therapy studies include:
If you are using Lifeline® cells to perform transfection and gene therapy experiments, let us know! Your studies could be featured