Human Pancreatic CAFS and Tumor Dynamics

 Extracellular Matrix (ECM) Matters
Our Human Pancreatic Cancer Fibroblasts were recently featured in a publication focusing on the role of the ECM on tumor dynamics. Andreas Stylianou, Vasiliki Gkretsi, Maria Louca, Lefteris C. Zacharia, and Triantafyllos Stylianopoulos (2019). Collagen content and extracellular matrix cause cytoskeletal remodelling in pancreatic fibroblasts. J. R. Soc. Interface 16: 20190226. http://dx.doi.org/10.1098/rsif.2019.0226.

Commercially available pancreatic native human FBs and,CAFs (cat. nos. SC00A5 and CAF08, respectively, Neuromics) were cultured in MSC-GRO (VitroPlus III, low serum, complete,
cat. no. SC00B1, Neuromics) medium in a 5% CO2-incubator at 37^OC.

Figure: Representative CAFs spheroids embedded in collagen gels at time 0 and at 6 h post implantation, respectively.

The results demonstrated that collagen concentration and consequently ECM stiffening differentially modulates FBs and CAFs cytoskeleton remodelling and invasion with stress fibre orientation being significantly altered in FBs. Finally, the presence of TGF-b reverses the suppressive effect of collagen stiffness on cell invasion.

3-D Tissue Models Made Easy

3 Steps to an in vivo Like Assay
We provide both CollaGel Hydrogels with different stiffness options and ready made 3-D Human Tissue Models.

 Building these models can be done in 3 easy steps:

• Step 1: Thaw our CollaGel Hydrogel solution 
• Step 2: Add your cell suspensions 
• Step 3: Watch it gel

HUVECS intergrated into 3-D CollaGel environment.

Due to the way in which it gels, our Collagel Hydrogel is well suited to Bio Printing models.

I will be posting more data as it becomes available.

Check Out Our 3D Human Skin Tissue Model

In vivo Like Model for Studying Skin Diseases/Irritants

I am pleased to announce this new addition to our Cell Based Assay offerings.

3D Human Skin Tissue Model

Cross section of Human Skin Model

The model includes precultured tissue in our engineered and tissue/cell specific Collagel-Hydrogels.

Our 3D human skin model is manufactured in a GMP lab. The model is engineered from multilayered, differentiated epidermal cells. The tissue is provided cultured in inserts using serum free media. The model mimics human skin. It provides an in vivo like environment for studying dermal related diseases and cancers. It is also excellent for skin toxicology and irritancy assays.

Check all our  3D Models:

Name	Applications	Size	Price
Human Blood Brain Barrier Model	In vivo Like Assays	6 wells 12 well 24 well	$1,780 $2,600 $3,625
3D HUVEC Model	In vivo Like Assays	6 well 12 well 24 Well	$1,269 $1,399 $1,539
3-D Human Skin Tissue Model	In vivo Like Assays	6 well 12 well 24 Well	$1,269 $1,399 $1,539

Questions? Contact me directly-Pete Shuster, CEO and Owner, 612-801-1007 or pshuster@neuromics.com.

3-D Screens Predict Breast Cancer Metastasis

Cell-ECM Interactions

3-D culturing solutions and methods are increasingly being used to created more in vivo like assays.

Here researchers use our Human Collagen IV extracellular matrix protein (ECM) to develop a simple biomaterial platform with systematic control over the ECM protein density and composition to determine if integrin binding governs how metastatic cells differentiate between secondary tissue sites: L.E. Barney, E.C. Dandley, L.E. Jansen, N.G. Reich, A.M. Mercurio, and S.R. Peyton. A cell–ECM screening method to predict breast cancer metastasis. Integr Biol (Camb). 2015 Feb 10; 7(2): 198–212. doi: 10.1039/c4ib00218k.

This publication details the creation of an in vitro fingerprint that is predictive of in vivo metastasis.

Figure: Biomaterial platform for integrin-mediated phenotyping. (a) Breast cancer cell lines with their known in vivo metastatic tropisms.  (b) Three distinct ECM microenvironments regulate integrin binding. (c) Adhesion and motility phenotypes of the MDA-MB-231 cell line. Black: ECM 1; blue: ECM 2; green: ECM 3.

Figure: Correlations between adhesion and migration responses identify potent integrin antibodies in vitro. Pairwise comparisons between adhesion and migration measurements in the (a, b) MDA-MB-231 and (c, d) SkBr3 cell lines across normal, EGF-stimulated, and integrin antibody conditions. Arrows highlight conditions where integrin antibodies increased migration metrics. Spearman correlations are indicated on each plot with two-tailed p-values. Circle: ECM 1; square: ECM 2; triangle: ECM 3; black: normal; green: EGF; blue: anti-β1 integrin; red: anti-α2 integrin; orange: anti-α6 integrin. (e, f) SkBr3 migration mechanisms are displayed via 10 random cell paths under (e) normal and (f) anti-β1 conditions. Red paths identify cells detaching and adhering elsewhere on the surface. Inset: representative images of cell morphology. Scale bar is 25 μm. (g) Individual cells that invaded into an overlaid 3D collagen gel from the ECM 1 surface after 48 hours. Bar indicates mean distance invaded of all invading cells. Inset: schematic of cells invading upward from the ECM surface into an overlaid gel.

A richer understanding of cancer metastasis is important for the development of new, more potent chemotherapies. I am hopeful 3-D/ECM assays become more embraced as a way to more accuratly determine in vivo cell behaviors.

Neurons in 3-D

In Vivo Like Neurite Outgrowth Cultures

Culturing in 3-D requires potent Primary Neurons. Here researchers use our e18 Rat Cortical Neurons in developing their 3-D assays: Chandrasekhar R. Kothapallia, Peyman Honarmandib. Theoretical and experimental quantification of the role of diffusive chemogradients on neuritogenesis within 3D collagen scaffolds. Acta Biomaterialia. Available online 14 May 2014. http://dx.doi.org/10.1016/j.actbio.2014.05.009

Abstract: A critical challenge to regenerating close mimics of native axonal pathways under chronic neurodegenerative disease or injury conditions is the inability to stimulate, sustain and steer neurite outgrowth over a long distance, till they reach their intended targets. Understanding neurite outgrowth necessitates quantitative determination of the role of molecular gradients on growth cone navigation under dynamic physiological conditions. High-fidelity biomimetic platforms are needed to computationally and experimentally acquire and analyze spatio-temporal molecular gradient evolution and the growth cone response across multiple conditions along this gradient pathway. In this study, we utilized a simple microfluidic platform in which diffusive gradients were generated within a 3D porous scaffold in a defined and reproducible manner, and its characteristics (spatio-temporal gradient, steepness, diffusion time, etc.) precisely quantified at every specific location within the scaffold. Using this platform, we show that the cortical neurite response within 3D collagen scaffolds, at both the cellular and molecular level, is extremely sensitive to subtle changes in localized concentration and gradient steepness of IGF-1 within that region. This platform could also be used to study other biological processes such as morphogenesis and cancer metastasis, where chemogradients are expected to significantly regulate the outcomes. Results from this study might be of tremendous use in designing biomaterial scaffolds for neural tissue engineering, axonal pathway regeneration under injury or disease, and in formulating targeted drug delivery strategies.

Image: Neurons in 3-D Assay

Neuromics' provides many Stem and Primary Cell Assay Solutions including tools for 3-D Cultures. We also offer services studying the effects of small molecules and compounds on Stem Cell expansion, differentiation and migration. To learn more contact me at 612-801-1007 or pshuster@neuromics.com.