Neuroscience Cells, CAFS, Cancer Cells plus BBB and HUVEC Models

Yvonne Adams, Anja T.R. Jensen (University of Copenhagen) manufactured Organioids using Neuromics' Primary Human Astrocytes and Human Brain Microvascular Endothelial Cells to study Plasmodium falciparum erythrocyte membrane protein 1 variants induce cell swelling and disrupt the blood–brain barrier in cerebral malaria.

Cerebral malaria (CM) is caused by the binding of Plasmodium falciparum–infected erythrocytes (IEs) to the brain microvasculature, leading to inflammation, vessel occlusion, and cerebral swelling.

Image 1: 3D BBB spheroids are composed of three different cell types. (A) FACS histogram showing pericytes (NG2, neural/glial antigen 2), astrocytes (GFAP, glial fibrillary acidic protein), and human cerebral microvascular endothelial cells (hCMEC/D3 and CD31).

Image2: ICAM-1–enriched microvilli and transmigratory ring/docking structures on brain hCMEC/D3 endothelial cells. (A–C) hCMEC/D3 brain endothelial cells were incubated with parasites.

This is first time the presence of intact, mature P. falciparum IEs within brain microvascular endothelial cells both in vitro and in vivo.  The data on the enhanced binding and internalization of IEs suggest that the same approach will be necessary to counteract not only the effects of cytoadhesion but also the subsequent contribution to potentially lethal brain swelling in CM.

With colorectal cancer (CRC) being one of the most prevalent cancers worldwide, understanding chemotherapy resistance in the CRC tumor microenvironment (TME) is an important goal. As we’ve seen in many other publications using our human cancer-associated fibroblasts (CAFs), CAFs play a huge role in the TME.

Our CAFs have been used in a handful of studies in 2D and 3D TME models, where they demonstrate their ability to complicate treatment objectives. Overall, the mechanisms by which CAFs contribute to chemotherapy resistance in the TME are poorly understood. 

Image: DKK1 expression in Neuromics colorectal CAFs when subjected to chemotherapeutic agents.

Researchers from the University of Kentucky utilized our colorectal CAFs (cat. CAF115) in a new paper examining these mechanisms. In their cell models, they found that CAFs secrete DKK1, a protein associated with tumor growth and metastasis, when subjected to chemotherapy treatments for CRC. Their results identified CAF-secreted DKK1 as a major player in resistance to treatment. Furthermore, they recommend looking into treatments that target DKK1 to counteract chemotherapy resistance.

You can check out the full paper here.

When researchers employ our human primary cancer associated fibroblasts (CAFs) in their tumor microenvironment models, they consistently find stark differences between 2D and 3D. Through 3D co-culture, investigators recognize that their models mirror in vivo tumors more closely than in 2D.

This makes our CAFs invaluable, enabling drug discovery research that better mimics the tumor microenvironment. Furthermore, over the last few years, we've seen the observation across many of our CAF types, including pancreatic, colorectal, breast, and now, ovarian.

Image: Human ovarian cancer cell lines (SKOV3 and OVACR8) along with Neuromics ovarian CAFs (cat. CAF112) in 2D and 3D ovarian cancer models.

Just how important is the tumor microenvironment (TME) in cancer research? As a supplier of primary human cancer-associated fibroblasts (CAFs), we’re exposed to plenty of studies looking into their role. One big theme: it’s becoming ever clearer that cancer treatments must take CAFs and their role in the TME into account.

Image: 3D organotypic colorectal tumor model surrounded by CAFs (vimentin, green) (a). Organoids are larger and more plentiful when CAFs are incorporated than without CAFs (b & c).

In a newly released paper, a team of researchers from the University of Akron builds 3D organoids with patient-derived colorectal tumor samples to study prospective cancer treatments. They compare how organoids develop, prosper, and react to treatments when they are with and without colorectal tumor CAFs (cat. CAF115) provided by Neuromics.

In this study, the authors developed a 3D functional human microvascular network in a microfluidic device. The established model enables Neuromics GFP-labeled human umbilical vein endothelial cellsto form vessel-like microtissues and have physiological functions which are closer to cells in human blood vessels. The perfusable microvasculature allows the delivery of nutrients, and oxygen, as well as flow-induced mechanical stimuli into the luminal space of the endothelium. The microflow effectively mimic the blood flow in human vessels.

This in-vivo like model is then used for toxicity assays-Yan Li, Qing-Meng Pi, Peng-Cheng Wang, Lie-Ju Liu, Zheng-Gang Han,Yang Shao, Ying Zhai, Zheng-Yu Zuo, Zhi-Yong Gong, Xu Yang and Yang Wu. Functional human 3D microvascular networks on a chip to study the procoagulant effects of ambient fine particulate matter. : RSC Adv., 2017, 7, 56108

Our human primary and stem cells are widely used and frequently published. We will continue to post relevant results from researchers using the cells here.

Images: Microvascular network formation based on microfluidic 3D HUVEC culture. (A) Schematic diagram of a microfluidic device. (B) Schematic diagram of microvascular network formation based on microfluidic 3D HUVEC culture. (C) Schematic diagram of loading microparticles in microvascular networks. (D) Microscope image of HUVECs seeding in fibrin hydrogel. (E) Confocal microscope image of fluorescent microvascular networks

As always, Neuromics greatly appreciates when our products are used in publications. The most recent publication shared by customers took advantage of our mouse Tuj-1 antibody (Cat.#MO15013). In this publication, researchers describe how they created an ex vivo 3D vascularized neural constructs that mimics the function of the blood-brain barrier.

Image: The neurovascular interface containing Tuj1+ (green) neurons differentiated from NSCs (in spaces surrounding the vasculature network) and Claudin-5+ (red) endothelial cells (inside the microfluidic channels). Scale bar, 500 μm. Image citation below

Citation:

Haibing Yue, Kai Xie, Xianglin Ji, Bingzhe Xu, Chong Wang, and Peng Shi. (2020). Vascularized Neural Constructs for Ex-Vivo Reconstitution of Blood-Brain Barrier Function. Biomaterials. doi: 10.1016/j.biomaterials.2020.119980

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In addition to our Tuj-1 antibody used in this publication, Neuromics offers a wide selection of antibodies ideal for BBB research. Check them out here!

Image: Spatial distribution of neurons (Tuj1+, green) in the 3D neural construct mimicking the in vivo BBB. The gray dash lines indicated the interfaces of collagen and tubal networks. Citation same at the above image