Human Cancer Associated Fibroblasts

Customer Generated Data
Our Cancer Associated Fibroblasts (CAFs) are widely used and frequently published.

We always welcome customer data. We are especially pleased when the data confirms our cells are "walking and talking" as advertised. Here's a recent example, "Biomarkers expression by flow and confocal microscopy using your CAFs. As flow cytometry indicated, >92% cells were live and we showed expected signals (Cy3) from the live cells. Nuclear dye is Hoechst showed as blue." Courtesy of Jiehua Zhou, City of Hope.

We offer a wide range of human primary cells. Check them out today.

Sleeping Beauty (SB) Transposon for the Study of Cancers

SB Gene Insertion vs Viral Based Methods

With the addition of SB Transposon Systems to our tool set. We have deep interest in how well SB works vs alternative methods. Here Dr. Hyun-Pyo Kim and team demonstrate how the SB Transposon system can be used for Novel Therapeutic Approaches for Various Cancer Types Using a Modified Sleeping Beauty-Based Gene Delivery System (DOI: 10.1371/journal.pone.0086324).

SB vs Viral Based Methods: "Successful gene therapy largely depends on the selective introduction of therapeutic genes into the appropriate target cancer cells. One of the most effective and promising approaches for targeting tumor tissue during gene delivery is the use of viral vectors, which allow for high efficiency gene delivery. However, the use of viral vectors is not without risks and safety concerns, such as toxicities, a host immune response towards the viral antigens or potential viral recombination into the host's chromosome; these risks limit the clinical application of viral vectors. The Sleeping Beauty (SB) transposon-based system is an attractive, non-viral alternative to viral delivery systems. SB may be less immunogenic than the viral vector system due to its lack of viral sequences. The SB-based gene delivery system can stably integrate into the host cell genome to produce the therapeutic gene product over the lifetime of a cell. However, when compared to viral vectors, the non-viral SB-based gene delivery system still has limited therapeutic efficacy due to the lack of long-lasting gene expression potential and tumor cell specific gene transfer ability. These limitations could be overcome by modifying the SB system through the introduction of the hTERT promoter and the SV40 enhancer. In this study, a modified SB delivery system, under control of the hTERT promoter in conjunction with the SV40 enhancer, was able to successfully transfer the suicide gene (HSV-TK) into multiple types of cancer cells. The modified SB transfected cancer cells exhibited a significantly increased cancer cell specific death rate. These data suggest that our modified SB-based gene delivery system can be used as a safe and efficient tool for cancer cell specific therapeutic gene transfer and stable long-term expression."

Figure 8. The effect of modified SB system on the tumor growth in vivo. Lung cancer cells (H358) (A), prostate cancer cell line (DU-145) (B), and ovarian cancer cells (OVCAR3) (C) were harvested by trypsinization, and 1×105 viable cells (as determined by trypan blue exclusion) in a total volume of 200 µl were injected subcutaneously. Two days following tumor seeding, animals were intravenously injected via tail veins with 100 mg/kg gancyclovir (GCV) along with either co-transfection of the empty plasmid (pT. hTp. Con) with the active helper plasmid (pCMV-SB) or co-transfection of the SB system (pT.hTp.HSV-tk.Con) with the active helper plasmid (pCMV-SB). Mice were sacrificed 28 days after tumor injection, and the effect of modified SB system on tumor growth was evaluated by measuring tumor size. doi:10.1371/journal.pone.0086324.g008

Neuromics, with our partner B-Mogen, has the capabilities to engineer custom SB Transposons for your Cancer Research. If interested, please contact me directly at 612-801-1007 or pshuster@neuromics.com. Our process is to first completely understand your unique requirements and from these, formulate a related statement of work with costs, timeline, milestones and deliverables. Thank you. Pete Shuster, CEO and Owner,

Cancer Research-Why 3-D Cultures Work

The concept of bringing your cancer cell assay to life with 3-D nanofibers is new. New approaches beg the question, "why change?". Traditional 2-D cultures are the standard and work well enough for many assays.

Change is driven by proof that the new solutions yield better results. I would like to share data showing the potential capabilities and benefits of switching to random and aligned nanofibers.

Increased drug sensitivity

Increased drug sensitivity of human A549 lung cancer cells when grown on random nanofibers or aligned nanofibers when compared to flat tissue culture plastic (TCP). This shows the significant problem of developing drugs using 2-D surfaces and explains why such a large amount of animal testing is required for pre-clinical drug development. Proliferation on 2-D TCPS is artificially high when compared to 3-D culture on nanofibers. Using a high-throughput 3-D nanofiber-based scaffold for in vitro drug screening can more accurately predict the in vivo response of drugs.

The ability to do high-resolution imaging through the nanofiber scaffold (through the bottom of the culture plate) is critical to validate cell/phenotype markers especially in high throughput screening and high content analysis.  Standard microscopes and automated plate readers using light, fluorescence, absorbance, or luminescence are compatible with the nanofiber plates.  Comparison of A549 cells on flat tissue culture polystyrene (A), randomly oriented nanofibers (B), and aligned nanofibers (C).

Cancer Cell Migration-More Like in vivo

Scanning electron microscope images of an ex-vivo human glioblastoma tumor sample cultured on aligned nanofibers showing the tumor dispersion along the nanofibers exactly how they would migrate in vivo along the white matter within the brain and central nervous system.

I will continue to post new developments with our 3-D culturing products. It is our goal to bring your cell based assays to life!