Biogelx are happy to welcome on board Africa Galvez Flores, who has just joined the Biogelx team as our resident cell biology researcher. Africa will be working on the development of realistic 3D cancer models using Biogelx proprietary peptide hydrogel technology. These models will be used to test the delivery and effectiveness of bio-orthogonal catalytic systems for the treatment of cancer as part of the Theracat Project.
Previously, Africa worked on neurodegeneration research at University College London Institute of Neurology and Columbia University Medical Center in New York, studying molecular aspects of neuronal death in cellular models of Parkinson’s and Alzheimer’s. She holds an MSc Neuroscience from University College London and a BSc Biochemistry from the University of Navarra. Nowadays, she is pursuing a scientific career in industry, focused on the usage of innovative approaches to treat disease.
Africa will be working alongside the rest of the Biogelx team in Biocity, Scotland, and will also be trained on novel cancer therapies in close collaboration with the rest of the Theracat consortium partners. Read more about this fascinating project that we’re just kicking off with Africa, here.
The aim of the THERACAT network is to consolidate Europe as the world leader in novel catalysis-based approach for cancer therapy.
David Hughes is one of our new Biogelx sponsored PhD students.His MRS funded project aims to develop a reproducible and easily manipulated 3D osteoarthritis in vitro model by using BiogelxTM peptide hydrogelsto examine bone and cartilage crosstalk in healthy and pathological ageing. In this interview, he speaks about his research and shares his hands-on experiences working with our products.
Please tell us a little bit about yourself.
I’m a PhD student from central Scotland with a keen interest in biomedical science; I originally undertook an undergraduate degree in Biology and Psychology. Due to my interest particularly in the biological side of my studies I then went on to pursue an MSc in Biomedical Science and Drug Design. This masters gave me an insight into tissue culture and its potential research value which lead me to where I am today. Outside of my studies I am a keen guitarist and play as part of a band.
What research do you focus on?
My research is focused on the development of a novel 3D model to investigate the crosstalk between bone and cartilage in osteoarthritis (OA). Currently there is no validated in vitromodel of the whole joint for OA research and the limitations posed by current 2D techniques necessitates the development of an appropriate 3D model that is more like the in vivoconditions. My PhD is funded by Medical Research Scotland in the lab of Dr Katherine Staines at Edinburgh Napier University.
Why is it so important to use synthetic 3D cell culture materials in your research?
Synthetic 3D culture materials such as those produced by Biogelx, provide a reliable and reproducible platform that allows me to conduct a variety of techniques on my cells of interest. Importantly synthetic 3D culture systems help eliminate further factors introduced through the variability associated with the use of a biologic system.
Have you tried any alternative materials? What are the key features of a good 3D cell culture?
I have not tried any additional 3D culture materials; however, I believe that the key features of a good 3D cell culture system are biocompatibility, modifiability and ease of handling. These features can allow for the investigation of a broad spectrum of variables across a variety of cell types whilst allowing the researcher additional optimization options which can save time and provide financial benefits.
How easy is to use Biogelx products?
Biogelx products are very easy to use, it’s as simple as weighing a powder out, rehydrating it and then adding the cell culture media that contains divalent cations to promote cross-linkage. The resources required to do this are commonplace in biomedical laboratories, keeping the gel preparation process simple and accessible.
What cell types do you use in your research? Please share your experience in growing cells in our gel(s). If you have any pictures that you can share, please do.
My research is still in early stages – I’ve been working with chondrocytes in the gels and have had no issues so far. Chondrocytes are the cells found in the cartilage, which covers the ends of the bones in the joint. Preliminary imaging shows that the cells seeded on the gels take on the characteristic rounded chondrocyte shape associated with their in vivomorphology; chondrocytes plated in a monolayer do not display this and take on a fibroblast-like shape, which is not ideal for investigating these cells and their functions in vitro.
What are the next steps in your research?
The next steps in my research will involve characterising the morphology and molecular phenotype of the chondrocytes and comparing it between cells cultured in monolayers (2D) and in Biogelx hydrogels (3D). Following on from this I intend to conduct similar optimisation on cells from the other joint tissues, before looking to develop my 3D model which I will use to investigate the crosstalk between bone and cartilage in OA.
3D bioprinting is a highly promising technique in the field of medicine. The technology is being used to in a range of 3D cell culture applications, from research projects developing bioprinted 3D tumor models, which better mimic the actual tumor environment, to projects which aim to create realistic 3D tissue constructs with the potential to implant them to the human body in future clinical applications. Whilst the concept of printing body parts here on Earth may seem far-fetched enough to some, and in reality is still a while away, there are some researchers taking this concept to the next level, and considering the clinical applications of bioprinting in space.
A Study on the Survivability and Adaptation of Humans to Long-Duration Exploratory Missions (HUMEX) discusses the potential health-related issues including survivability and adaptation of astronauts undertaking long-duration missions. The study highlights the effects of cosmic radiation, reduced gravity on Moon and Mars, gravity changes during launch, and related psychological health issues. Let’s imagine a one hundred and eighty days stay on the Moon with a crew size of four. Getting medical attention to the traveling crew is nearly impossible. Communication of medical instructions via IT and telecommunications technology (telemedicine), will be difficult due to the communication delay on deep space missions. In the event of a medical emergency, a rapid return home will not be feasible. Therefore, the patients will have to be treated on the spot. To find a solution for this issue, scientists are evaluating the feasibility of technologies such as 3D bioprinting in future exploration missions.
Doing 3D bioprinting in space, like on Earth, requires careful control of the extrusion process and control of the machine’s operating temperatures. However, the microgravity environment lacks the convection we are accustomed to here on Earth. Hence, we have to develop specific printing systems and bioinks which are able to perform in a microgravity environment.
To alleviate the issues caused by the specific space environment, a strategic partnership has been developed between the bioprinter company Allevi and Made in Space. The strategic alliance has designed a printer, which is capable of printing biomaterials in microgravity. The project aims to place this machine aboard the International Space Station (ISS), and could be considered the first step toward those scenes of sci-fi medicine.
There is also a project lead by the European Space Agency (ESA), which intends to advance 3D bioprinting to the level where it becomes possible to 3D print skin, bones and organs.
The human organ has a complex network of cells, tissues, nerves, and structures. They need to be positioned with the highest precision so that the organ functions properly. Currently, we do not have the technology to precisely blueprint these complex networks using different materials, cell types and bio-inks in the same final structure. Companies like Organovo are working hard to meet this challenge.
Although the printer technology and printing processes will be key to bioprinting in space, the biggest hurdle may still be the materials. Although the printer technology and printing processes will be key in developing bioprinting technology for space travel, another hurdle will still be the materials. In the future, if a medical emergency arises on a space mission where bioprinting could provide an immediate solution, access to reproducible bioinks with high biocompatibility and consistent printability could be vital.
An article by MijoSimunovic and colleagues from the Brivanlou and Siggia labs at The Rockefeller University in New York City showcases the use of Biogelx hydrogels in a 3D model of a human epiblast.
The authors used human embryonic stem cells (hECSs) to create an in vitro 3D model of an epiblast in which size, cell polarity, and gene expression mimicked those of a 10-day human epiblast.
The epiblast is a polarized epithelium, which will give rise to all cells of the fetus. During gastrulation, the three body axes (anterior-posterior, dorsal-ventral and the left-right axes) are created, establishing the vertebrate body plan. Mouse gastrulation 3D models (called gastruloids) have been previously generated from mouse embryonic stem cells (mESCs), and the morphogenetic movements that establish body symmetry at gastrulation and much of the signaling network underlying these processes have been elucidated in the mouse. However, some of these complex mechanisms are different in humans. Moreover, directly studying human gastrulation and human embryonic development is very challenging due to ethical guidelines (among other considerations) and has been largely unexplored. Therefore, synthetic models of human embryos are needed to address these questions.
In this work, the authors initallly used Matrigel to culture 3D hESC colonies, however observed the cells differentiated despite the absence of added morphogens. Thus, the researchers concluded that they could not use a Matrigel system to model a pre-gastrulating human epiblast. To improve culture conditions, Simunovic and colleagues utilized Biogelx peptide hydrogel as the 3D matrix.They observed how the 3D hESC colonies displayed “…similar size range, morphology, molecular markers, and tissue polarity that matched our in vitro attached 10-day human embryos”. Furthermore, cells cultured in this 3D model formed a quasi-spherical hollow shell and uniformly expressed key biomarkers like NANOG, SOX2, and OCT4 unlike in pure Matrigel. Additional parameters like the radius of the human epiblast and the surface cell density compared very well with their model epiblast.
This shows how Biogelx, a peptide hydrogel with defined composition and controlled chemical and mechanical properties proves to be very useful in modelling complex systems which are aim to elucidate intricate processes. Biogelx gels can be utilized either combined with other biomaterials or on their own; especially when using the new functionalized products which are based on the original Biogelx technology, but also contain specific biomimetic sequences relevant to the ECM and basement membrane proteins.
REFERENCES: Simunovic et al. (2018) Molecular mechanism of symmetry breaking in a 3D model of a human epiblast.bioRxiv 330704.
Extracellular matrix (ECM) mimics are used in a wide range of applications in cell culture, tissue engineering, regenerative medicine and the fast-growing market of bioprinting. Like the ECM itself, which is the non-cellular component present within all tissues and organs, they provide structural, biological and mechanical support which allows cells to survive, grow, migrate and differentiate and help maintain normal homeostasis. Often such ECM mimics come the form of hydrogels, and can be either naturally-derived or synthetic, with both having theirs benefits.
Naturally-derived materials have inherent biological properties which can more closely mimic the in vivo cell environment, often providing cell adhesion sites and great biocompatibility for cells. Hydrogel materials such as collagen or the complex protein mixture Matrigel are widely used in research, and have allowed the generation of a multitude of invaluable cell culture data as a result. There can be drawbacks associated with the use of these naturally-derived ECM mimics however, as batch to batch consistency is renowned for being variable which can be troublesome for sensitive or long term experiments. Often, temperature dependency/sensitivity of these materials can cause practical issues for users e.g. having to chill equipment and work on ice to maintain an optimum temperature can impart added complexities to experimental procedures.
Synthetic ECM mimics are increasingly being used as they can provide solutions to some of the issues that naturally-derived materials pose. The main benefit of synthetic hydrogels is consistency. There should be no batch to batch variations as they are produced using known quantities, optimised procedure, and generally the chemistry of the polymer does not change unless required. As they are synthetic, there are no components from animal-derived sources which can be an important factor for cell culture and tissue engineering research with the ultimate aim of clinical applicability. Synthetic ECM mimics also have the advantage of being carefully manipulated to suit an individual application. This can be done by tailoring the physical properties, but also by controlling the chemical structure of the materials. For example, the ability to incorporate biomimetic peptide sequences such as the RGD motif from fibronectin , or GFOGER from collagen at carefully controlled concentrations can be crucial for many synthetic matrices to provide good cell recognition sites. Indeed, with the addition of such components at similar concentrations to those found in vivo, synthetic ECM mimics have the potential to rival their naturally derived counterparts.
The ability to combine the benefits of both naturally-derived and synthetic hydrogel matrices could provide a product which would mimic the biological, mechanical and structural support found in native ECMs, but with batch to batch consistency, easy handling and the ability to manipulate. A totally synthetic ECM containing a combination of biomimetic peptides sequences would provide the user with a more consistent product, whilst still retaining biologically active sites found in protein-based matrices.
Biogelx currently provides 5 off the shelf synthetic hydrogel products including 4 containing biomimetic peptide sequences from ECM proteins fibronectin, laminin, and collagen (RGD, IKVAV, YIGSR and GFOGER respectively), each demonstrating benefits to different cell types. We are excited to share that we have been developing our next generation of peptide hydrogel product! In this single product we combine such biomimetic sequences in ratios which match the protein composition of a highly effective naturally-derived hydrogel product, and thus more closely mimic the natural ECM. With this new product, we hope to bring users the best of both worlds, providing the biological relevance of naturally-derived materials with the consistency of a synthetic hydrogel. Watch this space for more info and details on the lauch!