Roslin Cellab Ltd

Roslin, United Kingdom

Roslin Cellab Ltd

Roslin, United Kingdom
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Faulkner-Jones A.,Heriot - Watt University | Fyfe C.,Roslin Cellab Ltd | Cornelissen D.-J.,Heriot - Watt University | Gardner J.,Roslin Cellab Ltd | And 5 more authors.
Biofabrication | Year: 2015

We report the first investigation into the bioprinting of human induced pluripotent stem cells (hiPSCs), their response to a valve-based printing process as well as their post-printing differentiation into hepatocyte-like cells (HLCs). HLCs differentiated from both hiPSCs and human embryonic stem cells (hESCs) sources were bioprinted and examined for the presence of hepatic markers to further validate the compatibility of the valve-based bioprinting process with fragile cell transfer. Examined cells were positive for nuclear factor 4 alpha and were demonstrated to secrete albumin and have morphology that was also found to be similar to that of hepatocytes. Both hESC and hiPSC lines were tested for post-printing viability and pluripotency and were found to have negligible difference in terms of viability and pluripotency between the printed and non-printed cells. hESC-derived HLCs were 3D printed using alginate hydrogel matrix and tested for viability and albumin secretion during the remaining differentiation and were found to be hepatic in nature. 3D printed with 40-layer of HLC-containing alginate structures reached peak albumin secretion at day 21 of the differentiation protocol. This work demonstrates that the valve-based printing process is gentle enough to print human pluripotent stem cells (hPSCs) (both hESCs and hiPSCs) while either maintaining their pluripotency or directing their differentiation into specific lineages. The ability to bioprint hPSCs will pave the way for producing organs or tissues on demand from patient specific cells which could be used for animal-free drug development and personalized medicine. © 2015 IOP Publishing Ltd.


Faulkner-Jones A.,Heriot - Watt University | Greenhough S.,Roslin Cellab Ltd. | A King J.,Roslin Cellab Ltd. | Gardner J.,Roslin Cellab Ltd. | And 2 more authors.
Biofabrication | Year: 2013

In recent years, the use of a simple inkjet technology for cell printing has triggered tremendous interest and established the field of biofabrication. A key challenge has been the development of printing processes which are both controllable and less harmful, in order to preserve cell and tissue viability and functions. Here, we report on the development of a valve-based cell printer that has been validated to print highly viable cells in programmable patterns from two different bio-inks with independent control of the volume of each droplet (with a lower limit of 2 nL or fewer than five cells per droplet). Human ESCs were used to make spheroids by overprinting two opposing gradients of bio-ink; one of hESCs in medium and the other of medium alone. The resulting array of uniform sized droplets with a gradient of cell concentrations was inverted to allow cells to aggregate and form spheroids via gravity. The resulting aggregates have controllable and repeatable sizes, and consequently they can be made to order for specific applications. Spheroids with between 5 and 140 dissociated cells resulted in spheroids of 0.25-0.6 mm diameter. This work demonstrates that the valve-based printing process is gentle enough to maintain stem cell viability, accurate enough to produce spheroids of uniform size, and that printed cells maintain their pluripotency. This study includes the first analysis of the response of human embryonic stem cells to the printing process using this valve-based printing setup. © 2013 IOP Publishing Ltd.


Grant
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: HEALTH.2013.0-1 | Award Amount: 5.23M | Year: 2014

Cell-based screenings are today a necessary tool for all types of clinical development and/or market approval of new drugs and chemicals. The major change in the last decade is a shift towards more physiologically relevant yet complex and sensitive cell models, like stem cells, and more recently, the shift to human induced pluripotent cells. Stem-cell technology has the potential to revolutionize drug discovery, making models available for primary screens, secondary pharmacology, safety pharmacology, metabolic profiling and toxicity evaluation. The overall aim of DropTech is the development of automated handling processes for stem cells with integrated readout methods, required for the use of stem cells in high-throughput assays such as the embryonic stem cell test (EST). DropTech will result on the one hand in a fully automated screening platform, usable by the industrial partners for reproducible and standardized high-throughput screening services and aggregate production. On the other hand a system that is directly exploitable for industrialization and marketing will be available to perform reliable and fast at least semi-automated screening approaches for customers. Therefore, the complete workflow of the EST, including stem cell expansion, embryoid body formation in hanging drops and transfer to 2D conditions will be automated using robotic and microfluidics systems. This will enable standardized, fast and efficient embryotoxicity screenings reducing the need for animal tests. DropTech will enable testing in a small- and medium-scaled budget accessible for SMEs and academia in the field of biotech and biomedicine. The DropTech platform will have therefore a significant impact on the development of new medication and therapies and will enable personalized medicine approaches as well as - in future - regenerative medicine. DropTech facilitate the use of cell models with highest biological relevance (human pluripotent stem cells) in their native conformation.


Greenhough S.,University of Edinburgh | Greenhough S.,Roslin Cellab Ltd | Bradburn H.,Roslin Cellab Ltd | Gardner J.,Roslin Cellab Ltd | Hay D.C.,University of Edinburgh
Cellular Reprogramming | Year: 2013

We have devised an embryoid body-based screening method for the selection of human embryonic stem cell (hESC) lines capable of forming functional hepatocyte-like cells (HLCs) after single-cell dissociation. The screening method highlighted one cell line from a panel of five that produced albumin-positive cells during embryoid body (EB) formation. Cell lines that did not produce albumin-positive cells during EB formation were shown to respond less well to directed differentiation following single-cell replating. Additionally, the seeding density of the pluripotent populations prior to differentiation was shown to exert a significant effect on the hepatic function of the final population of cells. In summary, we have developed a simple procedure that facilitates the identification of human hESC lines that tolerate single-cell replating and are capable of differentiating to HLCs. Although the hepatic function of cells produced by this method is ∼10-fold lower than our current gold standard stem cell-derived models, we believe that these findings represent an incremental step toward producing HLCs at scale. © 2013, Mary Ann Liebert, Inc.


Grant
Agency: GTR | Branch: Innovate UK | Program: | Phase: Feasibility Study | Award Amount: 117.43K | Year: 2013

Human liver hepatocytes are extremely valuable cells both for drug development laboratories and for patients whose own livers have become damaged either through viral infections, poor diet and/or long term excessive alcohol consumption. There is already a shortage of donor livers for transplantation and this is set to worsen in the future as liver failure rates continue to increase. It is known that stem cells can be multiplied greatly in culture and changed into hepatocytes to give an endless supply. However, the liver also has other cell types in it which contribute to efficient liver function. In this project we will identify these cell types and their ideal ratios to liver hepatocytes. This should form the basis for further development work towards recreating functional liver tissue from human stem cells.

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