News

Branco et al. have recently published an excellent paper showing the benefits of Atelerix technology in the preservation of Adipose-Derived Mesenchymal Stromal Cells. Demonstrating the maintenance of MSC viability and therapeutic function, they conclude Atelerix technology is a viable solution for the storage and distribution of cell therapy products.

Access the full article here.

A big congratulations to all the authors who have worked hard on this paper, it is a great read!

MSc André Branco

MSc Ana L. Tiago

Dr. Paula Laranjeira

MSc Maria C. Carreira

MD João C. Milhano

Dr. Francisco dos Santos

Prof. Joaquim M. S. Cabral

MD Artur Paiva

Prof. Cláudia L. da Silva

Dr. Ana Fernandes‐Platzgummer

How preservation of urine can impact kidney disease research

Background:

Each day, 20 people in the UK alone, develop chronic kidney disease, while an estimated 3 million people in the UK are thought to suffer from some form of kidney disease. Kidney disease can occur due to 1) genetics, 2) poorly controlled diabetes, 3) heart disease, and 4) obesity. When the kidneys stop working properly, harmful toxins and excess fluid can build up in the body. People suffering from kidney disease also may suffer from high blood pressure, anaemia and bone and muscle weakness.

Primary ciliopathies are a group of rare genetic conditions, which can affect multiple organ systems including the kidneys (Figure 1). Found in almost all human cells, primary cilia are finger-like protrusions, acting like radio antennae receiving information from the surrounding environment. The primary cilia are also responsible for key cellular signalling mechanisms. Defects in these primary cilia are responsible for primary ciliopathies.

Figure 1. Wheel depicting primary ciliopathies which are known to have a renal phenotype. Other hallmark characteristics of each ciliopathy are also shown, to highlight the broad spectrum and organ-system involvement associated with each primary ciliopathy. Figure generated using BioRender.com

How we currently study primary ciliopathies:

To study primary ciliopathies, we use primary ciliated cells. One of the most frequently used cell lines to do this is fibroblasts, often derived from skin biopsies. Researchers often chose to ship established, confluent fibroblasts live, in flasks full to the brim with growth media. Another method of sample shipment commonly used is cryopreservation. However, delays in transport during long-haul shipments pose a major threat to sample integrity. Evaporation of dry ice, and consequently thawing of samples, results in cells sitting in dimethyl sulphate (DMSO), which is toxic at ambient temperatures.

Another rapidly growing method to study renal ciliopathies focuses on the use of urine. Every day, between 2000 and 7000 cells are shed from the kidneys, into the urine. These cells, known as human urine-derived renal epithelial cells (hURECs) can be isolated to study kidney diseases on a patient-specific basis. We use these hURECs to study several disease aspects, including primary cilia phenotype (how the cilia look and behave), investigating potential genetic drivers of specific diseases, and nephrotoxic drug screening (Figure 2). Currently, as researchers, we must collect, transport, and process urine samples within 4-hours, on ice, to ensure that these cells are viable for expansion. This is not an issue for locally based patients, however, for those patients further afield this 4-hour time frame poses a major hurdle. To complicate matters further, as hURECs are a primary cell line, they tend to be more sensitive to traditional cryopreservation shipment methods; in fact, long-distance collaborating researchers often opt to send hURECs live in plates (increasing contamination risks and likelihood of cell death).

Figure 2. Overview of how human urine-derived renal epithelial cells (hURECs) are currently isolated from whole urine samples, and how these cells are utilised in kidney and ciliopathy-focused research. Figure generated using Biorender.com.

How Atelerix technology can help:

Atelerix offers a solution to this sample shipment problem. Their novel technology specialises in the preservation of biological material at room temperature, removing the need for cell exposure to harmful cryopreservation reagents (like DMSO), eliminating the need for dry ice, and removing the risk of sending live cells. It is hoped that the specialist hydrogel preservation technology developed by Atelerix could be implemented at one of three key time points in the processing of urine samples (Figure 3).

Figure 3. Overview of the urine processing sample, with areas where Atelerix products could be implemented to help overcome issues associated with long-distance samples shipping shown in purple boxes. Areas: 1) after spinning down urine samples, Atelerix products (BeadReady or CytoStor) could be utilised to preserve the whole urinary pellet (which contains hURECs). 2) After urine spinning, and resuspension of the whole urinary pellet, cell suspension could be plated, and the Atelerix WellReady kit used to preserve samples prior to shipment. 3) Following complete urine processing, established hURECs could be shipped using any of Atelerix’s Kits (BeadReady, CytoStor or WellReady). Figure generated using BioRender.com.

Next Steps:

Over a three-month period, the following two aims will be explored to try and establish the impact of Atelerix technologies on renal and primary ciliopathy research.

1. Explore the possibilities of using Atelerix technologies to preserve cells during the early stages of hUREC isolation
2. Increase the viability of preserved established ciliated cell lines (including primary hURECs and fibroblasts) following preservation at room temperature to mimic shipment periods

Becky Dewhurst

MRC DiMeN DTP iCASE PhD Student

Newcastle University

Pancreatic cancer is one of the most aggressive and lethal solid tumours. Pancreatic
ductal adenocarcinoma (PDAC) describes malignancies of the exocrine pancreatic
tissue, that account for 90% of all pancreatic cancer cases. Unfortunately, PDAC is a
highly metastatic cancer that is often discovered at the end-stage of the disease. At
this point, conventional treatments, such as surgical resection and chemotherapies
mostly fail. This highlights the need for personalised drug testing platforms, that
allow to quickly evaluate the most promising combination of chemotherapeutic
agents or novel therapies for the individual PDAC patient. To that end, preserving
primary PDAC clinical samples (e.g., biopsies and surgically resected tissues) is
crucial, as these patient tissues can be used for establishing patient-specific tumour
organoid models to be used in screening for treatments.

Figure 1. From the clinic to the lab: primary patient tissue samples are used for patient-derived organoid modelling. Surgically resected pancreatic tumour tissues or patient biopsy samples need to be preserved during transport from the clinic to the lab. At the lab, the primary patient tissues are used for establishing in vitro models, such as patient-derived tumour organoids, that can used for screening most efficacious cancer treatment for the individual patient.

When the biological samples from cancer patients obtained in the clinic need to be transported to the lab for further research or personalised screening of therapies, conventionally, the samples are frozen, or cryopreserved. Yet, the cycles of freezing and thawing can damage the architecture of the tissues we aim to maintain. Interestingly, encapsulation within an algae-derived gel – alginate – could potentially be used as an alternative to freezing. Samples preserved within alginate are easier to handle, whether by car or via mail, as they can be shipped at ambient temperatures and there is no need for specialised vehicles or storage freezers. However, it is still unclear whether the ever adapting and shapeshifting cancer cells and tissues can be reliably stored within the alginate gel.

In vivo, the tissue-specific extracellular matrix (ECM) surrounding the cells naturally contains ligands that the cells can bind for anchorage, via integrin receptors.  Integrin-mediated cell-ECM interactions play a key role in cell survival, proliferation, differentiation, and migration. The plant-derived alginate matrix lacks the respective bioactive ligands required for integrin-mediated cell attachment to matrix. That is, for mammalian cells, alginate is biologically inert. It is known that the lack of anchorage to the matrix and contact inhibition from the increase of cell-to-cell interactions can result in cell cycle arrest. I.e., cells should stop growing and proliferating in alginate. However, contact inhibition is one of the anticancer mechanisms that is often lost in malignant cells.  This begs the question: can PDAC samples be reliably stored in alginate? – Will they slumber? Or will the cancer cells continue growing and thriving regardless?

To understand the response to PDAC cultures and tissues to alginate encapsulation, I am carrying out my LifETIME CDT PhD Bioengineering project at the University of Glasgow and CRUK Beatson Cancer Research Institute, supervised by Prof Laura Machesky, Prof Huabing Yin and Dr David Chang. With the help of each of the scientists bringing in their unique expertise on pancreatic cancer, material engineering and clinical practice respectively, I will be able to characterise the biological response of both in vitro PDAC models and patient clinical samples to short-term storage within alginate and examine the effects of the physical alginate properties on sample preservation.

Using both the user-friendly Atelerix Ready-alginate kits and own in-lab alginate, conventional PDAC cell lines, three-dimensionally cultured tumour organoid models and primary cancer tissues will be encapsulated and stored in the hydrogel. Cell survival and behaviour in alginate will be characterised using foundational cell read-outs like viability and morphology as well as quantification of gene and protein expression (e.g., measuring the levels of cell cycle genes). Metabolomic screens will also be performed to better understand PDAC sample phenotype and activity throughout storage within alginate. To probe whether the material characteristics of alginate play in PDAC sample preservation efficacy, elasticity and viscosity of alginate will be characterised. It is yet unclear whether the physical properties of alginate have any effect on sample over the short-term, e.g., 5-day, storage. However, since in vivo PDAC cells experience a stiffened surrounding (fibrotic) matrix and respond to mechanical signals, it is key to examine how the stiffness of alginate used for encapsulation may affect the preservation of PDAC cultures and tissues.

Figure 2. Cells stored in alginate do not directly adhere to the surrounding matrix. When pancreatic cancer tissue sample is encapsulated within alginate, the cells do not form connections to the matrix. Due to the lack of cell-matrix interactions, there is an emphasis on cell-to-cell contacts. However, it is yet unclear whether physical characteristics of the alginate gel, such as viscosity and elasticity matter for the short-term storage of pancreatic cancer tissues and cells in alginate.

In short, this project will help illuminate whether solid tumour models and primary patient tissues can be successfully preserved at ambient temperatures using alginate, and the extent to which physical alginate properties matter for this purpose. Since alginate preservation is low cost and does not require specialised equipment, it would allow more researchers to make use of tumour tissues and engage in patient tissue-derived organoid modelling. Overall, reliable storage within alginate would help to optimise sample preservation from the clinic to the lab bench, from one lab to another, strengthening the networking between the clinical, academic, and industrial scientists by easing the transport and sharing of advanced in vitro models and primary patient tissues.

Juda Milvidaite, PhD student (they/them)
EPSRC-SFI CDT in Engineered Tissues for Discovery, Industry and Medicine (LifETIME)

CRUK Beatson Institute

University of Glasgow

The population of haematopoietic stem cells (HSCs) found in our bone marrow are responsible for repopulating the body with all blood and immune cells throughout our lives. As we age, our HSCs can collect DNA mutations, and this can result in many blood disorders like leukaemia. Methods of curing blood cancers require HSC’s for either the testing of drugs and treatments, or physically replacing faulty HSCs with those from someone else, known as a stem cell transplant (SCT, commonly called a bone marrow transplant).
Unfortunately, the availability of donor HSCs is limited because, out of the body, in 2D cultures they lose their ‘stemness’; the ability to become dormant (quiescent) for longevity and self-renew for population maintenance, and instead differentiate; this means that they cannot engraft and renew the blood supply in a cancer patient. Yet vast quantities are required in the development of a drug (multiple rounds of drug screening), and around 2 million are required  per Kg of body mass for SCT.
Therefore, there is great need for in vitro methodologies that sustain this most primitive group and the project I am undertaking stems from this issue.

Firstly, I intend to form a 3D model of the most common place for HSCs to reside – the bone marrow (BM) microenvironment coined the ‘BM Niche’. The aim of any in vitro model is to replicate an environment in vivo. In the case of the bone marrow, it has been shown that marrow HSCs can be maintained by utilising a hydrogel scaffold* to mimic the physical features of the niche (stiffness), while biological additives such as extracellular matrix proteins and growth factors (structural and signalling proteins of the bone marrow), and cellular co-culture with marrow mesenchymal stromal cells (MSCs)** mimic the biochemical features that help support the HSCs.
Secondly, one type of signalling molecule secreted by MSCs to help support HSCs are bioactive
metabolites*** which signal to HSCs and result in retained stemness. Using this niche model, I shall isolate these metabolites and add them artificially to a simplified niche models without any biological additives with the aim of ‘tricking’ the cells into thinking they are in an optimal
environment and so that they retain their HSC characteristics for long enough to allow drug
screening.

This is where Atelerix come in, as their alginate hydrogels induce cell quiescence, enabling short-term storage of various cell types from days to weeks. We are looking to take advantage of this feature using their bead-ready kits, to enable the encapsulation, storage, and distribution of the core components of the simplified ‘niche’ model – HSCs, MSCs and metabolites. We are then left with a neat little package with the basic ‘niche’ components required for HSC maintenance that can be kept at room temperature and without media changes for long enough to be sent and delivered to labs around the world, who themselves are now able to grow their own population of stemness maintained HSCs or use the niches for drug testing directly without needing to thaw any cells.

To do this I am initially testing the viability of MSCs arranged into round clusters of cells, termed spheroids encapsulated in bead-ready kits. MSCs are being used initially as they retain primitive characteristics (especially when in spheroid formation and when encapsulated in a soft hydrogel) for much longer than HSCs when grown in 2D. The end product will consist of 96 ‘mini niches’ within a 96 well plate. To do this, we aim to use a bioprinter to print well-ready encapsulated cells and metabolites in a highly controlled and reproducible environment, ensuring batch-to-batch consistency not only between each niche, but between each 96 well plate of niches. This ensures the product is a viable option for drug testing as variance between niches and therefore testing conditions is minimised.

In summary, haematopoietic disorders lack in treatments due to the cost and poor availability of primary human HSCs. This project should yield a simplified niche model to grow human HSCs
without loss of stemness, the core components of which shall be encapsulated and stored within Atelerix ‘bead-ready’ gels. This will improve the availability of HSCs and in turn should greatly enhance the development of treatments for blood disorders and reduce the cost of stem cell treatments.

*Hydrogels have high water content, collagen, one of the most abundant proteins in our body forms hydrogels and this gives structure to our soft tissues and is why hydration helps these tissues.

**Mesenchymal stromal cells are a stem cell containing population that help repair, bone, cartilage,
ligament, and tendon. However, in the bone marrow, they spend most of their time looking after
HSCs, telling the blood stem cells what to do.

***Metabolites are biological small molecules made by cells to and which drive metabolism
(respiration, energy supply, amino acid synthesis (amino acids are the building blocks of proteins and
so help in making of tissues).

The EPSRC-SFI Joint Centre for Doctoral Training in Engineered Tissues for Discovery, Industry and Medicine (lifETIME CDT) is a partnership between the University of Glasgow, University of Birmingham, Aston University and CÚRAM – Science Foundation Ireland at the University of Galway.

The lifETIME CDT will train innovation leaders in drug discovery and regenerative medicine through development of bioengineered humanised 3D models, microfluidics, diagnostics and sensing platforms, focusing on developing technologies that replace and reduce use of animals in research.

Research students within lifETIME complete a 4-year PhD and undertake a range of skills training designed to help them network and develop leadership skills. The cohort-based training will forge a UK community of over 80 talented researchers with high value skills sought by the market and who can deliver change.

In partnership with industry, the lifETIME CDT co-create 4-year PhD projects to address industry specific challenges. In collaboration the lifETIME CDT, Atelerix have co-created two research projects and work together with lifETIME academics to support and supervisor the research students. The lifETIME CDT highly values the partnership with Atelerix, and the opportunities this creates to deliver enhanced training for our students.

The CDT students work with Atelerix on projects where room temperature storage of cells 3D models would be an advantage to make cold chain simpler or the envisaged tissue easier to use.

If you would like to learn more about lifETIME CDT please contact Matthew Dalby (Matthew.Dalby@glasgow.ac.uk) and Michelle Carmichael (Michelle.Carmichael@glasgow.ac.uk)

One of the biggest challenges faced by CAR-T Cell manufactures is the variability among apheresis product, which can negatively impact T Cell manufacturing success and therefore patient treatment. A solution is required to minimise this variability such that the CAR-T manufacturing process can be standardised in accordance with Good Manufacturing Practices (GMP). We show that our technology, LeukoStor™ T25, enhances the predictability of starting T Cell yield compared to freeze-thaw and no-gel controls (Fig 1a). LeukoStor™ 25⁰C maintains >90% T-Cell viability after 5 days’ storage (Fig 1b), providing a 2-fold improvement compared to unprotected material (no-gel control). In apheresis material, the expected T Cell proportion is 53.8±6.1% (1), which is maintained by LeukoStor™ after 5 days’ storage (Fig 1c). Assessment of viability and metabolic capability following overnight culture shows that cells also maintain post-rest function after storage in LeukoStor™ (Fig 1 d, e).

The benefits of LeukoStor™ T25 address one of the critical drivers of CAR-T manufacturing delay by demonstrating how this cell storage technology can provide consistent, high quality starting material, offering a step towards both standardization and a reduction in vein-to-vein time (V2V). The current 2–3-week V2V time, defined as the time that elapses between apheresis and product delivery at the hospital, can be problematic and impact patient eligibility given the rapid progression of the disease. Implementation of LeukoStor™ T25 may offer a reduction in V2V and in turn reduced cost, through optimising time-limiting quality control processes or even storing cells from early apheresis. For CAR-T developers, complex cold chain also remains a challenge, for the transport of apheresed and manufactured cells between manufacturing facilities and hospitals. LeukoStor™ offers the potential to mitigate the need for temperature-controlled transport, through successful cell storage at 25⁰C.

References

1. Blood [Internet]. miltenyibiotec. [cited 2022 Sep 12]. Available from: https://www.miltenyibiotec.com/GB-en/resources/macs-handbook/human-cells-and-organs/human-cell-sources/blood-human.html

Atelerix’s hydrogel-encapsulation technology TissueReady™ has previously
demonstrated fantastic results at preservation of fresh tissue. The existing product is
tissue and user friendly and requires no formalin fixation. Currently we are further
refining TissueReady™ specifically for the purpose of extending the shelf life of fresh
human biopsies to enable improved diagnostics and patient monitoring in clinical
trials and in pre-clinical development. The aim is to allow tissue storage and
shipment for longer periods. This will improve scheduling demands, geographic
distribution and allow centralised testing of clinical biopsies ultimately positively
impacting users both financially and in terms of the quality of service they provide.
To date, an initial assessment of product performance has been performed with
encouraging results. Human breast, colorectal and skin tumour tissue was stored
successfully in the refined TissueReady™ formulation with no detrimental changes to
the structure or decrease in viability of the tissues after the 96-hour storage period.
Next steps are to undertake full histopathological and functional studies on stored
cancer tissue as well as design a tissue storage solution compatible with large tissue
resections (up to 30cm). We hope that this will improve the downstream assessment of
tumour samples, for example removing the challenges of obtaining genomic material
from FFPE samples to perform NGS and allow research and clinical professionals to use
fresh human tissue samples in diagnostics, drug discovery and clinical indications.

New distributor agreement between Atelerix (UK) and BioBench Solutions LLP (India).

We are proud to announce the beginning of a new distribution agreement with BioBench Solutions, who will be distributing our products across Indian.

Our global customer base is growing rapidly due to problems associated with cryogenics, the vast increase in the use of complex physiologically relevant cell models in drug discovery, and our movement towards the regulated market of cell and gene therapy. Under this agreement, BioBench Solutions will increase our visibility on the Indian Market and distribute Atelerix’s portfolio of hypothermic cell, tissue, and virus preservation technology.

About BioBench Solutions

Biobench Solutions, headquartered in Pune India was founded in 2017 to distribute high quality and innovative products within research and diagnostics. Given our great experience and knowledge, our sales and technical support teams have the laboratory experience needed to provide high quality support to customers regardless of research discipline.

We are proud that our proactive approach to development in response to our customers’ needs has become one of our strengths and has led to our future-oriented foresight and deep trust. We continue to work together in creating better future where people can live in good health, both physically and mentally.

biobenchsolutions.com

European Society of Gene and Cell Therapy Conference

29^th Congress

11^th – 14^th  October 2022

Edinburgh, Scotland

Booth P5 – 01

Email lindsay.smith@atelerix.co.uk to book a meeting

www.esgctcongress.com

We are excited to announce the release of the newest addition in our hypothermic cell and virus preservation range.

CytoStor’s cytoprotective capacity can encapsulate up to 10 million cells per tube, making it the most scalable solution we have brought to market. Our new streamlined and automatable application ensures users can encapsulate their cells and viruses more easily than ever ensuring all your needs from research through to production are met.

It is as simple as mixing your cells with our specially formulated hydrogel solution, transferring the solution into the storage vial and you are done!
If you are new to our hypothermic preservation technology, take a look below at the effects on cell preservation for Human Adipose-derived Mesenchymal Stromal Cells, Hela Cells and Human airway epithelial cells.

Atelerix is looking to recruit two research and development scientists to help drive our product development. We’re looking for an Immunologist and a Histologist to expand the capabilities of our team. These essential roles within the R&D team will support the design, development and validation of products with a range of biologicals including different cell types, 3D culture models, and primary tissues.

Research and Development Scientist – Immunology

Essential Duties and Responsibilities:

Design and execution of experimental workplans that focus on the preservation of blood products and isolated immune cells. Knowledge of molecular and cellular biology, and specifically immune cell biology, is essential.

Duties include:

• Processing different types of human blood products (e.g. LRS cones, leukopaks & peripheral whole blood), including isolation of PBMC and immune cell subsets
• Phenotyping subsets of immune cells by performing immunolabelling and flow cytometry.
• Analysing flow cytometry data
• Handling and assaying the phenotype and function of lymphocytes, monocytes and macrophages
• Reviewing, collating, evaluating relevant literature
Writing and reviewing protocols
• Sourcing of cells/consumables
• Project planning
• Performing and writing risk assessments
• Conducting practical experiments
• Troubleshooting
• Data analysis and report writing
• Internal / external dissemination of results

Product Development

Design and execution of experimental workplans on generating new product formats to solve existing problems (e.g. integration with different cell culture formats & manufacturing processes)

• Product design
• Product evaluation
• Protocol construction

Essential Requirements:

• A PhD in a biological science discipline
• Extensive cell culture experience and/or handling primary cells/tissues
• Familiarity with a range of experimental and analytical techniques
• Experience in developing, optimising and executing biological assays, ideally in a commercial setting.
• Excellent written and verbal communication skills
• Hardworking and self-motivated
• Excellent problem-solving skills
• Legal right to work and reside in the UK

Desirable:

• Experience in culture and assay of primary blood cells/immune cells
• Experience with cell biochemical and metabolic assays
• Experience of working within a QMS (Quality Management System) framework
• Previous employment in the pharmaceutical or biotechnology industry would be an advantage
• Experience of working with industry partners and stakeholders
• Experience of grant writing and application processes

Apply Here

Research and Development Scientist – Oncology/Histology

Essential Duties and Responsibilities:

Design and execution of experimental workplans that focus on the preservation of solid tissue biopsies. General knowledge of molecular and cellular biology is required; specific knowledge of tumour histopathology would be highly desirable.

Duties include:

• Handling and processing solid tissue samples for histology
• Performing standard (e.g. H&E staining) and specialised (e.g. IHC) histological techniques
• Viewing, imaging, and analysing histological data in an accurate and reliable manner
• Writing and reviewing protocols
• Sourcing of cells/consumables
• Project planning
• Carrying out and writing risk assessments
• Conducting practical experiments
• Troubleshooting
• Data analysis and report writing
• Internal / external dissemination of results

Product Development

Design and execution of experimental workplans on generating new product formats to solve existing problems (e.g. integration with different formats & manufacturing processes).

• Product design
• Product evaluation
• Protocol construction

Essential Requirements:

• A PhD in biological science discipline
• Extensive cell culture experience and/or handling primary cells/tissues/biopsies
• Familiarity with a range of experimental and analytical techniques, particularly associated with tumour histology
• Experience in developing, optimising and executing biological assays ideally in a commercial setting
• Excellent written and verbal communication skills
• Hardworking and self-motivated
• Excellent problem-solving skills
• Legal right to work and reside in the UK

Desirable:

• Significant experience of experimentation with tumour biopsies across the entire workflow: from collection & handling to processing and characterisation
• Knowledge of pathology, particularly cancer pathology
• Experience with cell biochemical and metabolic assays
• Experience of working within a QMS framework
• Working with industry partners / stakeholders, preferably from within industry itself
• Experience of grant writing and applications processes

About Atelerix Ltd
Atelerix is a UK-based company whose patented technology is transforming the storage and transport of living cells by enabling extended storage at temperatures between 4°C and 25°C while preserving their functional viability. The company’s customers are developing cell-based therapeutics, cell lines, organoids and assays for use in drug discovery and development. Atelerix products eliminate the need for complex ultra-low temperature logistics and potentially harmful freeze-thaw processes, providing greater quality, consistency of performance, flexibility, convenience and security as critical samples are shipped around the world.