Cancer Associated Fibroblasts (CAFS) Traction Dynamics and Remodeling in 3D Matrices

 Neuromics' Colorectal CAFS in Action

In a first of its kind study, the mechanical properties of our CAFS were determined. This is important because, understanding how these cell behave in vivo will help improve the efficacy of new therapies: Bashar Emon, Zhengwei Li, Md Saddam Hossain Joy, Umnia Doha, Farhad Kosari, and M Taher A Saif. (2020). A Novel Method for Sensor-Based Quantification of Single/Multi-Cellular Traction Dynamics and Remodeling in 3D Matrices. bioRxiv. doi: 10.1101/2020.09.24.311647

"Cells in vivo generate mechanical forces (traction) on surrounding 3D extra cellular matrix (ECM) and cells. Such traction and biochemical cues may remodel the matrix, e.g. increase stiffness, which in turn influences cell functions and forces. This dynamic reciprocity mediates development and tumorigenesis. Currently, there is no method available to directly quantify single cell traction and matrix remodeling in 3D. Here, we introduce a method to fulfil this long-standing need. We developed a high-resolution microfabricated sensor which hosts a 3D cell-ECM tissue formed by self-assembly. It measures cell forces and tissue-stiffness and can apply mechanical stimulation to the tissue. We measured single and multicellular force dynamics of fibroblasts (3T3), human colon (FET) and lung (A549) cancer cells and cancer associated fibroblasts (CAF05) with 1 nN resolution. Single cells show significant force fluctuations in 3D. FET/CAF co-culture system, mimicking cancer tumor microenvironment, increased tissue stiffness by 3 times within 24 hours."

Here's one of 10 videos illustrating these properties.

 CAFS are widely used and frequently published. Check them out today!

Sorting Cancer-Associated Fibroblasts (CAFs)

Featuring Our Colorectal CAFS
CAFs play a central role in the Tumor Microenvironment (TME). The TME has been identified as one of the driving factors of tumor progression and invasion. Inside this microenvironment, CAFs, a type of perpetually activated fibroblasts, have been implicated to have a strong tumor modulating effect and play a key role in areas such as drug resistance.

This makes CAFs a target for cancer therapies. The challenge is the TME is heterogeneous making it a challenge to derive homogeneously relevant populations for basic research and drug discovery. This new study uses our Colorectal CAFs to identify markers for isolating these populations. Martin Nurmik, Pit Ullmann, Fabien Rodriguez, Serge Haan, Elisabeth Letellier. In search of definitions: Cancer-associated fibroblasts and their markers. doi: 10.1002/ijc.32193.

Images: Expression of common markers in patient-derived fibroblasts. Immunofluorescent staining of primary colon cancer fibroblasts (Neuromics, #CAF05), reveals a heterogeneous expression pattern for both αSMA/ACTA2 (abcam #ab7817, 1/200) and FAP (Santa-Cruz Biotechnology #sc-65398, 1/200), while PDGFRα (abcam #ab61219, 1/200) expression in tested cells remains relatively homogenous. Nuclei of fibroblasts were stained using DAPI (DAPI Fluoromount-G® Mounting Medium). Image is representative of three independent biological experiments.Cells were imaged using a Zeiss LSM 510 Meta laser scanning confocal microscope (Carl Zeiss, Jena, Germany) with a Plan-Apochromat 63x/1.40.

The authors conclude when selecting for CAF populations using antibody-based methods such as FACS, it is essential that multiple surface markers are used in order to avoid any chance of introducing unintentional subtype bias. Other available surface markers such as PDGFRα/β work well here, as do more general fibroblast surface markers like Thy-1 cell surface antigen (CD90), provided that the cell population is also subjected to selection with negative markers,

Cancer Associated Fibroblasts (CAFs)-Flow Cytometry and Immunostaining Results

If they Walk and Talk Like CAFs, They Are!

We are experiencing growing demand for are CAFs. They are closely associated with primary tumor cells and participate in the neoplastic process. There is reciprocal communication between CAFs and tumor cells through paracrine effects of secreted growth factors, cytokines & chemokines from both fibroblasts, tumor cells and other tumor-associated cells.

Potential users are asking how well the cells are characterized. The purpose of this posting is to share key gene expression data:

CELL LINES	CD105	CD90	CD44	CD326	CD133	FAP
SC00A1 (hMSCs)	+	+	+	-	-	-
SC00A5 (pancreatic fibroblasts)	+	+	+	-	-	-
CAF05 (colorectal CAFs)	+	+	+	-	-	+
CAF07-AD (adenocarcinoma lung CAFs)	+	+	+	-	-	+
CAF08 (Pancreatic Stellate CAFs)	+	+	+	-	-	+

Table: expression of key proteins

Images: Human Lung Adenocarcinoma CAFs stained with Vimentin, aSMA, b-Catenin and DAPI (blue),

Cancer Associated Fibroblasts Gene Expression Data(pdf - 784Kb)-Flow Cytometry and Immunostaining Results.

Questions? Please contact me directly-Pete Shuster- CEO & Owner-pshuster@neuromics.com or 612-801-1007.