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!

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.

Neuromics New FBS-329/500 ml.

Premium Ultra-pure FBS!
Check it out today

Ultrapure FBS is manufactured, by our OEM partner Biowest USA. in a fully integrated environment from raw material collection to finished product. Our production facility is fully certified to ISO 9000 and ISO 13485. Ultrapure means:

• Lowest endotoxin level 0,1 EU/mL. 
• Triple 0,1 μm Filtered. 
• Free of Virus and Mycoplasma. 
• Tested Ideal for Sensitive Cells and Applications. 

This FBS can be used to culture all cell types including macrophages, cancer and cancer related cells, hybridomas and more. It works great for all our human primary cells and stem cells.

Human Cells-utopia!

Great for use a controls vs Differentiated iPSCs

We have a cornucopia of Human Primary Neuron, Astrocytes and Schwann Cells plus CAFS. Featured Assays:
Human Brain Pericytes used to study Guide Axon Guidance.
Human Pancreatic CAFS and Tumor Dynamics.

Axon guidance at the site of a cervical spinal cord injury in a rat model. (Ai) Schematic illustrating transplantation of scaffold into a C-4 hemisection. The injury cavity is shown prior to (ii) and immediately following (iii) transplantation. (Bi) Scaffold conditioned with flow exhibits viable GFP-labeled microvessels (green) (ii) and alignment of host axons (magenta) infiltrating the scaffold in the rostral-caudal direction (grey arrow). (C) Scaffold conditioned in static conditions showing disrupted alignment of both microvessels (ii) and host axons (iii). (D–F) Microvessel and axon plots showing alignment (D,E) and length (F). Scale bars, 1 mm (Aii,Aiii) and 50 μm (B,C). Data are presented as mean ± s.e.m. ***P < 0.001; statistical significance was calculated using Welch Two Sample t-test. White arrows denote proximity of axons with microvessels. Microvessel alignment values (n = 30), axon alignment values (n = 30), microvessel length values (n = 15), and axon length values (n = 15) are from single hydrogel samples per condition.

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

It is imperative that all our cells work as advertised. If your results do not meet expectations, we will run similar tests to make sure they walk and talk as they should.

We wish you and yours a Happy Holiday Season, Pete Shuster, CEO and Owner Direct phone: 612-801-1007 or pshuster@neuromics.com

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,

Data Trifecta-Cells, Media + FBS

Optimizing your Cell-Based Assays
Researchers need solid proof that the solutions the use will work in their assays. Given the time and effort it takes to culture human primary cells, it is important that the media and supplements used are optimized.

We engineer our cell media and supplements to work with our primary and stem cells. Its potency is tested every day in our cell cultures.

As further proof, we ask customers to share their data with us. We reward them by emailing a 50 USD Amazon gift card. I would like to recognize Emily Rodela from TGEN for sharing the Trifecta of Neuromics' CAFS grown in MSC-Gro Media supplemented with our FBS.

Image: Human CAFS-Pancreatic Stellates cultured in MSCGro™ supplemented with Neuromics' FBS - Fetal Bovine Serum. Courtesy of Emily Rodela, TGEN.

Cancer Associated Fibroblasts in Action

Publication Reference
We introduced our Cancer Associated Fibroblasts to researchers several years ago. The demand was strong from day one and continues to grow.

We are pleased to announce the first reference in a Cancer Research Publication: Zenobia D'Costa, Keaton Jones, Abul Azad, Ruud van Stiphout, Su Y Lim, Ana L Gomes, Paul Kinchesh, Sean C Smart, W. Gillies McKenna, Francesca M Buffa, Owen J. Sansom, Ruth J. Muschel, Eric O'Neill and Emmanouil Fokas. Gemcitabine-induced TIMP1 attenuates therapy response and promotes tumor growth and liver metastasis in pancreatic cancer. Published OnlineFirst August 1, 2017 doi: 10.1158/0008-5472.CAN-16-2833...Immortalized Pancreatic CAF-Stellate Cells were obtained from Neuromics, and cultured in VitroPlus III, low serum, complete (Neuromics)...

Image: Pancreatic CAF-Stellates stained with alphaSMA

We will continue to post new developments here.

Human Melanoma Cancer Associated Fibroblasts

More Hard to Find Human Cells
We are recognized for providing potent, pure and easy to culture CAFs and Immortalized Tumor Cells. We are pleased to announce the addition of Melanoma CAFS. Our introductory price is only 899 USD/1,000,000 Cells. They have been successfully passaged 10X.

.

Image: Human Melanoma Cells in Culture

Need CAFs or Tumor Cells? Just email me at pshuster@neuromics.com.

Need Cells From Diseased Donors?

Just Ask
Some of our customers have expressed the need for Cells that have the phenotype of a given disease. This is especially true for researchers doing drug discovery for neuro-diseases like Alzheimer's and Parkinson's.

Deriving "hard to find" cells from tissue is one of our cornerstones.

If you have need for any type of human cell(s), give us a shot. I can be reached at 612-801-1007 or pshuster@neuromics.com. Thank you. Pete Shuster,CEO and Owner, Neuromics.

Cancer-Associated Fibroblasts and Cancer Cell Migration

Highway for Cancer Metastasis

Cancer Associated Fibroblasts provide the Matrices  (ECMs) serve as cancer’s assistants as it spreads throughout the body.

Cancer cells (red) migrate on a CAF-derived extracellular matrix (green). VANDERBILT UNIVERSITY; BEGUM ERDOGAN, DONNA WEBB

Unlike other ECMs, which normally form dense meshes, these arrange into parallel bundles forming the highway for tumor cell migration (The Scientist-December 15, 2015).

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.

Racing into 2015

Thank you for a Great 2014!
Wow. I would like to thank our customers, collaborators and friends for helping us achieve unprecedented success in 2014.

We plan on racing hard into 2015. I would like to highlight our focus areas for the New Year.

Stem Cell Activating Solutions-We have been pilot testing solutions foir balanced immunity and stem cell vitality. Our clients have been mostly peopole with autoimmune related issues. After 6 months of therapy, the improvement in symptoms and test results, for many, has been stunning (see below).
We have developed migration and other assays to test these solutions in vitro using our UCB Derived hMesenchymal Stem Cells. We also use medical testing to prove and improve their therapeutic value. These assays and arrays are available for labs and clinics interested in testing stem cell related solutions. Here're representative data from our Quantibody Array testing program.


Stem and Primary Cell Based Assays-We will continue to add new solutions for your assays. These will include mixed culture that more closely mimic in vivo environments like mixed astroglial-neuron cultures. Below is our hN2^TM Human Neurons stained with one of our neuron markers.

Image: hN2 cells grown in culture for 4 days and stained with our chicken polyclonal to Neurofilament light or low molecular weight chain NF-L, a marker of neurons. Many of the differentiating cells show strong cytoplasmic and clearly fibrillar staining for NF-L. Blue stain is DAPI and reveals cell nuclei of some non neuronal cells in this culture.

• Here're some of our human cells we added in 2014:
  Motor Neurons-GFP Labeled-Potent and Easy to Culture Mouse Motor Neurons
• Human Endothelial Cells-New-Artery, Microvascular, Vein & Umbilical Cord Derived + Culture Media
• Cancer-Associated Fibroblasts (CAFs) and Cells-Solutions for studying tumor growth, angiogenesis and metastasis.

---

Check out and join my Stem Cell Clinical Trails Group on Linkedin (4500+members strong).
We wish you a rewarding 2015.

---

Pete Shuster-
CEO & Owner--pshuster@neuromics.com or 612-801-1007

---

The Growing Value of Cancer Associated Fibroblasts or CAFS

Balancing Supply and Demand

There is reciprocal communication between Cancer Associated Fibroblasts (CAFs) and tumor cells. They are now recognized as playing a key role in promoting the malignant process and blocking immune surveillance  They are becoming increasing important in Cancer Drug Discovery Programs.

Their growing value is making it harder for us to source resected tumors. We have been receiving advanced orders from clients who need to insure they have a stock. We have ordered tumors for the below CAFs and will be shipping to those who have place advance orders by the end of November.

Name	Catalog #	Type	Species	Applications	Size	Price
Human Colorectal Tumor CAFs Kit	CAF05	Human Primary CAFs	H	Cell Assays	1,000,000 cells	$799
Human Endometrial Adenocarcinoma CAFs	CAF01	Human Primary CAFs	H	Cell Assays	1,000,000 cells	$799
Human Ovarian Serous Tumor CAFs	CAF02	Human Primary CAFs	H	Cell Assays	1,000,000 cells	$799
Human Lung Adenocarcinoma CAFs	CAF07A	Human Primary CAFs	H	Cell Assays	1,000,000 cells	$799
Human Lung Squamous Cell Tumor CAFs	CAF07SC	Human Primary CAFs	H	Cell Assays	1,000,000 cells	$799
Human Immortalized Pancreatic CAF-Stellate Cells	CAF08	Human Primary CAFs	H	Cell Assays	500,000 Immortized Cells	$4,335
Human Pancreatic Tumor Cells Kit	PXPC3A1	Cells+Media Kit	H	Cell Assays	1,000,000 cells	$499
Conditioned Media for Ovarian Serous Tumor CAFS	CAF04	Cell Growth Media	H	Cell Assays	5 ml	$250
Conditioned Media from Endometrial CAFS	CAF03	Cell Growth Media	H	Cell Assays	5 ml	$250
VitroPlus III, low serum, complete	PC00B1-100	Cell Growth Media	H	Cell Assays	100 ml 500 ml	$65 $165

Image: Endometrial CAFs courtesy of Tiana Tonrey, Vitro Biopharma

Should have questions about availability and ship dates for any of these please call (612-801-1007) or e-mail: pshuster@neuromics.com. Thank you.
Pete Shuster, CEO, Neuromics

Pancreatic Cancer Tumor Cell Assays

Providing Pancreatic Tumor Cells or Related Custom Assays

We are pleased to announce the addition of Pancreatic Tumor Cells to our Cell Based Assay offerings. We are also generating a wealth of internal data with our Tumor Cells and Cancer Associated Fibroblasts. We have now opened up our capabilities to Cancer Researchers via our new CRO services. These included customized assays for studying toxicity using compounds/small molecules. Here's an example:

In addition, we can do kinetic, live cell screens using these cells plus our hMSC and CAFs. To learn more call 612 -801-1007 e-mail me: pshuster@neuromics.com. We stand ready to serve you in a way that fits your specific requirements.

In vivo Like Cell Based Assays!

Save 100 USD on our Potent, Pure and Easy to Culture Stem and Progenitor Cells

We continue to design in vivo like behaviors into our  Cell Based Assays.
Our goal is to enable you to make better research decisions earlier
in your discovery process.

These assays are engineered using our proven, potent and easy to culture eSC and Adult Human Stem Cells and Progenitors.

UCB Derived hMSC differentiated to Osteoblast. Note the calcium depositions.

***December Promotions-New Products-Save 100 USD+ 
on Primary and Stem Cells, Media, Markers and Growth Factors***

• Human Mesenchymal Stem Cells-Passage up to 12X-395 USD 
• Native Human Chondrocytes-HCS-449 USD
• Native Human Osteoblasts-449 USD
• Cancer-Associated Fibroblasts (CAFs)-499USD
• Save 70 USD-only 225 USD for new Neuron, Astroglial and Progenitor Markers

I'll be posting videos of our in vivo like assays with specific quantitative data here soon.