Editor’s Choice for Core-Shell Capsules
The Article "Alginate Core-Shell Capsules for 3D Cultivation of Adipose-Derived Mesenchymal Stem Cells”, published in February 2022, was recently selected as "Editor's Choice". The paper was part of the Special Issue “Material and Engineering-Based Approaches for Organoids” in the Open Access journal MDPI Bioengineering and was the first lead-authorship publication of Sabrina Nebel, doctoral candidate at the Institute of Cell and Tissue Culture Technologies.
Sabrina’s research is focused on mesenchymal stem cells (MSCs), progenitor cells of the connective tissue that can be found in almost all tissues, even in adults. These cells are easily available and can be used in a wide variety of cell therapies and in-vitro testing systems. However, since millions of cells are required for any application, ex-vivo cell expansion is inevitable. Cultivation in petri dishes in 2D is still considered the gold standard, despite evidence that 3D cultivation not only retains but can even improve the beneficial properties of the cells.
Part of Sabrina’s PhD project is to develop such 3D cultivation systems. In her publication, she presents an approach in which the cells are enclosed in so-called core-shell capsules. These capsules consist of an outer barrier made from alginate and an inner, liquid compartment. This core contains the stem cells which can then form direct cell-to-cell contacts just as in their physiological environment. The alginate shell serves as a protective layer from mechanical stress and allows for nutrient and waste diffusion.
“I like to joke that I am making ‘Stem Cell Bubble Tea’ in the lab, as my capsules are made the same way as the popping bubbles you can find in lots of these tea shops” she explains. They are of course not edible, but will hopefully be a valuable tool for stem cell expansion in the future, paving the way for novel therapies.
Talk at ÖGMBT Annual Meeting
Julia Moldaschl is a doctoral candidate at the Institute of Cell and Tissue Culture Technologies (ICTCT) at the Department of Biotechnology since September 2021. One of her most exciting achievements so far was her contribution to the 14th ÖGMBT Annual Meeting, in Vienna in September 2022. She gave a talk titled “Multilineage Differentiation of Juvenile Primary Bone Marrow-Derived Mesenchymal Stem Cells in 3D under Normoxic and Physioxic Conditions” in the section “Cell Based Assays, Therapies and Products”. This posed an ideal opportunity for Julia to share her experience with mesenchymal stem/stromal cells (MSCs) cultured and differentiated under physiological conditions.
In particular, her research focuses on the establishment of a MSC multilineage differentiation model under physiological conditions, which comprise scaffold-free 3D culture approaches, reduced oxygen conditions, that can also be considered as “Physioxia”, and the application of xeno- and serum-free media.
Instead of broadly used 2D cultivation systems, Julia cultures MSCs as three-dimensional aggregates (spheroids) in micropatterned multi-well plates. That culture system enables the formation of uniform spheroids, allows for up-scaling and has been shown to lead to increased stemness and enhanced differentiation capacity for certain lineages. Oxygen conditions of MSCs´ natural microenvironment are aimed to be resembled by providing physioxic culture conditions. As the third major parameter, Julia is only using xeno-free and serum-free media instead of Fetal Bovine Serum (FBS) supplemented media. This selection is associated with increased proliferation rates and the elimination of ethical problems and risks of pathogen transmission originating from FBS. The application of all three mentioned culture conditions aims to recapitulate the in vivo situation of the cells and therefore increases the relevance of data derived from in vitro models, particularly in the context of MSC-based therapies.
Poster Prize – ÖGMBT Workshop 2023
On 17th March 2023 Julia attended the ÖGMBT Workshop “Advanced Cell Culture Technologies” and was awarded the prize for the best poster. The workshop was organized by the ÖGMBT working group “Cell based assays, therapies and products” and took place directly at BOKU. During the workshop Julia joined lively discussions, exchanged experience in 3D cell culture technologies and connected with new colleagues. “It is such an enriching experience! I got helpful feedback on my work and so many questions that benefited my thesis writing”. On top of this exciting day, Julia won the poster prize for her poster entitled “3D Multilineage Differentiation Model of Mesenchymal Stem/Stromal Cells under Physiological Conditions”.
First time presenting at a conference
Karolina Peter, member of the Doctoral School Biomaterials and Biointerfaces, could engage many interested listeners with her scientific presentation at the JUNIOR EUROMAT conference (19-22 July, Coimbra, Portugal). The presentation, titled "Zooming inside spider silk" - Using synchrotron nanoXRD to compare the ultrastructure of spider silks." was very well received and motivated many of the audience to ask interested questions. There were research results presented, which were collected at the Institute of Physics and Materials Science (BOKU).
Moreover, this was the first conference for Karolina that was not held in a virtual setting, and her first oral presentation.In addition to numerous other interesting lectures, one could also participate in workshops such as Electron Microscopy or Horizon Europe. In particular, the contact with other young scientists was great at JUNIOR EUROMAT 2022 in Coimbra. All in all, a very successful experience, both personally and scientifically valuable.
Research at a large-scale facility
Everything started with a remote experiment at the end of 2020, to which the team from the Institute of Physics and Material Science (IPM) was unfortunately unable to go in person due to Covid-19. Then, in June 2022, the time had finally come, and it was possible to travel in person to the ESRF (European Synchrotron and Radiation Facility) in Grenoble, France. The proposal for an experiment on spider silk was accepted, which is reason to celebrate.
After extensive preparation of samples (single fibers from different spiders as well as microtome sections of the same), the goal was to learn as much as possible about the ultrastructure of spider silk using nanobeam X-ray scattering. Also, thin sections of spider silk, which allow to look at the differences in the structure in a locally resolved way. On site, the team from IPM was supported by the scientists at the beamline (ID13 nanofocus beamline) and accompanied during the experiment. In total, the experiment lasted for 5 days, and a very large amount of data was collected, all of which is now waiting to be evaluated and analyzed. The team consisted of Karolina Peter (main responsible for the experiment), her colleagues Leon Ploszczanski and Arno Frank. Even though after the experiment the need to "catch up" on sleep and recharge the batteries was large (the synchrotron is in operation 24/7), it was an unforgettable and exciting experience, which will soon also result in an interesting publication.
My year of 2022 together with BioMatInt - Johannes Stöckelmaier
When I joined BioMatInt in 2021, I didn’t expect the changes it would bring to the path of my PhD-project. While I needed the first months to get familiar with topic and literature, BioMatInt gave me the opportunity to visit and learn from colleagues and research institutes abroad, leading to valuable gains in knowledge.
While I boarded my flight from Vienna to Copenhagen on March 1st, I knew that I would get to meet one of the most renowned professors for computer simulation of intrinsically disordered proteins in Europe. However, I did not expect to also find a group of great colleagues and a progressive and beautiful city to live in. There is a lot to learn from the city of Copenhagen, especially how to build a top-notch biking infrastructure; the best I have ever seen in my life.
I was able to significantly progress my PhD project, specifically in regard to ensemble reweighting; a topic the Lindorff-Larsen research group has vast experience in. I was given time to think through the mathematical details of their method and to experiment with their data.
During my stay in Copenhagen, I learned:
* … how to use the BME-Reweighting tool.
* … the workflow of a RNA reweighting-example.
* … insight into methodology and mathematical background of the BME method.
* … insight into danish culture and a (little) bit of their language
After visiting Scandinavia in the northern part of Europe, I had the chance to join a one-week long seminar about disordered proteins at Institut d'Etudes Scientifiques de Cargèse. Located on the island of Corsica, France, it was an adventure to get there with in-between stops in Paris, Nice and Ajaccio. The seminar was hold at a very remote location directly on the shore of the Mediterranean Sea. I was able to meet more than 30 like-minded colleagues from Europe, the US and from Japan. Lectures and trainings where held by senior scientists from France, Germany and the United States.
While joining the seminar, I learned:
* … how to use an ensemble refinement method.
* … how to use Flexible-Meccano to generate conformational ensembles.
* … how to approach IDP phase-separation simulation.
I’m very glad that BioMatInt gave me opportunity to take part in those events and I look forward to an equally successful 2023.
ATTENDANCE GRANT – ESCMID/ASM CONFERENCE 2022
In October 2022, Goodness Osondu-Chuka won an attendance grant to participate in and present her Ph.D. work at the ESCMID/ASM Conference on Drug Development to Meet the Challenge of Antimicrobial Resistance (2022), which took place in Dublin, Ireland.
Goodness is a doctoral candidate of the BioMatInt at the Institute of Biologically Inspired Materials. In her project, she creates a three-dimensional biofilm model that replicates key aspects of the biofilms found in the lungs of individuals with cystic fibrosis, a chronic illness without a cure. Through this model, she intends to investigate and comprehend the protective functions of biofilms in persistent infections, particularly in the lungs of cystic fibrosis patients, that prevent effective antibiotic treatments.
She applied for this grant because it offered an excellent opportunity to share and gain more knowledge on various aspects of microbial biofilms as well as novel techniques and approaches for solving the problem of antimicrobial resistance and tolerance in chronic infections. The grant allowed Goodness to present her first results on extracting, reconstituting, and printing the extracellular polymers that provide the structural properties of cystic fibrosis biofilms.
The conference, which was focused on the challenges of developing new agents and solutions for antimicrobial resistance, started with a half-day boot camp designed to share the wealth of experience from the community. This was followed by two and a half days of high-quality symposia, roundtable discussions, keynote, and young investigator lectures. The meeting was designed to bring together researchers, academics, industry regulators, and funders to share experiences, collaborate and learn. In her own words, “Attending the conference was a fantastic chance to stay informed about current research, express my thoughts, and network effectively. I eagerly anticipate similar opportunities to attend and actively participate in future conferences”.
Testing single spidersilk fibers at various humidity conditions - Leon Ploszczanski
The ESEM has not been used for the last two years. I had to completely skip and clean the water cooling to get the brown algae out of the pipes. But the result is very satisfying, you can examine a fresh organic sample without much drying effects in environmental mode with water vapour at 400 Pascal pressure. Between the sample, cooled to 4 °C, and the electron beam outlet are water molecules that protect the sample from drying out and also guide the secondary electrons to the detector. In the pictures you can see the setup in the chamber of the ESEM and in the micrograph a vacuole filled with essential oil in the outer shell of a blood orange.
First lead authorship publication
On February 2023 the article “Hydrogels from TEMPO-Oxidized Nanofibrillated Cellulose Support In Vitro Cultivation of Encapsulated Human Mesenchymal Stem Cells” was published on the ACS Applied Bio Materials journal. This was the first lead authorship publication of Ilias Nikolits, doctoral candidate at the Institute of Cell and Tissue Culture Technologies (ICTCT) at the Department of Biotechnology of BOKU.
The aim of Ilias’ PhD project is on the development of hydrogels for 3D in vitro cultivation of human mesenchymal stem cells (MSCs). The material he is focusing on his research is cellulose, a highly abundant and inexpensive natural polymer, with high biocompatibility and tunable properties. The hydrogel scaffolds synthesized from cellulose are aiming to provide a 3D in vivo-like environment during cultivation of MSCs in vitro, compared to the gold standard 2D plastic adherent methods, by preserving the physiological characteristics and functions of the cells.
In this publication, Ilias shows that hydrogels synthesized from chemically functionalized cellulose nanofibers consist a very promising platform for in vitro cultivation of humans MSCs. Encapsulation of MSCs in those hydrogels mimics a 3D tissue-like environment and cells can perform their physiological functions and behavior.
“When people ask me what hydrogels are, I give them the example of Gummy Bears. So, I am basically putting stem cells in Gummy Bears made from wood powder. But is seems the cells really like it” -Ilias quotes.
Research stay abroad - Magnetically triggered drug nanocapsules in hydrogels
University of Siegen, Germany (February 2023)
In February 2023, Giacomo Chizzola spent a month at the University of Siegen, Germany, to expand his knowledge and abilities in polymer synthesis. Giacomo, a member of the Biomaterials and Biointerfaces Doctoral School, is a doctoral candidate at the Institute of Biologically Inspired Materials.
His research focuses on developing the building blocks – specialised nanoparticles, oligomers, and polymers – as well as the strategies to achieve release of encapsulated drug compounds via magnetothermal actuation. To this end, Giacomo visited the partner laboratories of Professor Holger Schönherr and Professor Ulrich Jonas at the University of Siegen, in order to learn the synthesis procedures of some state-of-the-art polymers.
One of the polymers he produced is poly(ethylene glycol)-block-poly(lactic acid) (PEG-b-PLA), which is obtained via ring opening polymerization of cis-lactide onto a PEG macroinitiator. The synthesis can be adjusted to yield PLA blocks of varying lengths, which in turn result in different assembly architectures of the block copolymer in water. A second polymer that Giacomo worked on is poly(acrylamide-co-acrylonitrile), which was chosen for its tunable thermoresponsive properties. To have the best control over the polymer growth and critical solution temperature, he learned how to perform a sophisticated polymerization reaction called reversible addition-fragmentation chain-transfer (RAFT) polymerization.
Giacomo’s guest research stay in Siegen was enrichening and successful, as he came back to Vienna with a few different polymers to further modify and test for their properties and performance. He expects to return to Siegen later in the year or next year to learn more and produce more polymers tailored specifically to his research goals. Once back in Vienna, Giacomo commented the following:
“What I truly enjoy about my research project and being part of the BioMatInt Doctoral School is being immersed in such an interdisciplinary environment. My stay in Siegen was, in many ways, a true masterclass in polymer synthesis, a field in which I had very limited practical experience beforehand”.