FAQs

The first step is to contact one of our Account Managers who will give you an overview of our assays. If you wish to proceed, you will be introduced to the Technical Team. The Technical Team will take you through the process of study design and generate a Scope of Work (SOW). The scope of work presents the details of the study including deliverables and estimated timescales. At this point the Account Manager will send the final SOW and Quotation for the Study. If the SOW and price are agreed, the project will be assigned a Study Manager, an experienced laboratory scientist who will run the study and be your point of contact throughout the study. At the end of the study, the Study Manager will collate the results and the report.

Absolutely! Study design is a joint endeavour between your scientist and the Study Director and Study Manager at Newcells to produce the best study to answer your questions. Our experienced scientist may be able to guide you to the best solution with input from the client at every stage. Similarly, we can customise the report to your requirements and format.

Yes, communication is very important. We will keep you informed as we progress. Before we send results, we will perform a QC of the data and results. Equally, the Study Manager is available for teleconferences to discuss any ongoing study.

Retinal organoids contain major retinal cell types found in vivo. The differentiation process follows the developmental timeline of retinogenesis in vivo and cell types are arranged in stratified layers of the neural retina.

Yes, the organoids are light responsive.

We have standardised the retinal organoid manufacturing process and introduced a quality management system to ensure the process meets strict criteria. A degree of variability is expected in any biological system; however, all our organoids undergo rigorous quality controls to ensure experimental reproducibility.  Only batches that have passed our quality control are released to our clients.

Yes, we have proprietary transcriptomic data from retinal organoids at day 150 and day 200 of differentiation.

Our off-the-shelf offering of organoids as a ready-to-use assay is at day 150 of differentiation but this can be customised for other time points to fit your requirement from day 60 to day 210.

Retinal organoid differentiation process closely follows the developmental timeline of retinogenesis in vivo. Therefore, various cell types are on a different developmental trajectory. Transcripts for retinal ganglion cells are most abundant at day 60 of the differentiation, however photoreceptors mature at later stages and beyond day 150.

Retinal organoids do not typically contain RPE. We offer a separate service for RPE, please contact us for more details.

We currently offer RPE cells as part of a service which can be tailored to various customer needs. We are planning on offering RPE cells as a product in the near future.

We currently do not co-culture RPE cells with the retinal organoids. However, we can carry out studies using retinal organoids and RPE cells as two separate models.

We are able to offer CRISPR-Cas9 generated iPSC lines specific for your project needs. We are also working towards having a bank of cell lines available.

We can generate retinal organoids or RPE cells from customer iPSCs bearing a mutation. If the iPSCs are not available, they can be derived from patient cells or generated using CRISPR-Cas9 gene editing. We offer both reprogramming and gene editing service.

We recommend to use our wild-type (WT) qualified iPSC line to make any genetic modifications.

Generation of CRISPR-Cas9 iPSC lines takes 12-30 weeks depending on the editing complexity required. Generation of retinal organoids takes an additional 26 weeks for day 150 organoids.

We develop in vitro retinal assays for various applications and do not have capabilities for vector design. However, we have experience in working with gene therapy vectors through multiple customer and collaboration projects using our models.

We do not offer individual neural retinal cell types as a stand-alone product. However, we are able to provide scRNAseq of retinal organoids as a service to allow analysing separate cell populations, please contact us to discuss your specific project needs.

Standard retinal organoids are sold at day 150 of the differentiation, however depending on the application and project requirement they can be used at later stages. Earlier stages organoids can be kept for longer periods. Please contact us for more information.

Yes. Please refer to our recent publication where we assessed a broad range of transgene and capsid variants of AAV vectors. McClements et al. Transl Vis Sci Technol. 2022 Apr 1;11(4):3. doi: 10.1167/tvst.11.4.3.

Transduction efficiency is difficult to quantity in a complex multicellular model such as the organoids, and it varies depending on the AAV serotypes. In a collaborative paper, transduction efficiency has been evaluated by co staining of cell specific markers and transgene and transduction efficiencies of up to 37% have been achieved.

Mc Clements et al,(2022)  Transl Vis Sci Technol.;11(4):3 https://tvst.arvojournals.org/article.aspx?articleid=2778725

We can provide a RPE model which can potentially mimic early and dry forms of AMD.

The organoids are generated from iPSCs that were reprogrammed from human dermal fibroblasts obtained from healthy donors who provided full consent following formal ethical approval processes.

We are currently able to ship live retinal organoids to Europe and the USA.

Retinal organoids are also available as frozen pellets ready for RNA extraction or as frozen sections for immunostaining worldwide.

Retinal organoids are shipped live in 5 ml tubes containing 10 organoids each. In addition, we provide 150 ml of optimised cell culture medium and 2 x 96 well plates per 100 organoids, plus 2 x Pasteur pipettes for transferring them.

We have validated our shipment process for up to 7 days in transit, which is standard for all of our North American shipments.

All our organoid batches sold at day 150 of the differentiation have passed our quality control. In addition, our shipment processes have been validated. If you have any concerns, one of our technical experts will be able to provide you specific advice.

Yes, we can ship these to you.

In vitro assays such as our podocyte and proximal tubules models are well-qualified near physiological models of tissues that provide robust, reproducible data from which informed decisions can be made either about the value of in vivo studies in animals or to provide an understanding of unexpected liability at first in man. Furthermore, our models give a full picture of nephrotoxicity as we have a glomerulus and a proximal tubule model.

Our fresh proximal tubule model is available for shipment in some regions, but you can also outsource the work to us. Our glomerulus model is currently only available as part of a service.

Yes, you can, we will design the study with you and can provide you with either a data summary or a full report.

Yes, this is one of the major advantages of our model, we generate proximal tubule cells from mouse, rat, dog, human and even non-human primate. This allows accurate prediction in humans.

They are very predictive. For example, Takeda has done a study showing the high level of predictivity of our PTC model (Bajaj et al., 2020)

https://www.sciencedirect.com/science/article/abs/pii/S0300483X20301748

The cells are kept in a stasis media and require revival up receipt. They are shipped at 4 °C.

• Our fresh primary model is extremely well differentiated and representative of in vivo proximal tubule
• Our flagship model, aProximate™ is supported by a large package of validation data to support its use in transporter and nephrotoxicity assays and its value in predicting in vivo outcome. It is available across the main preclinical species (rat, dog, NHP and mouse) together with human to allow comparative studies.
• It has been used in regulatory submissions to support NDA and IND submissions
• If you choose to outsource to us, the service is performed in state of the art laboratories by experienced scientists working to strict SOPs and a Quality Management System.

The proximal tubules are arranged as a monolayer and express high levels of transporters than any other in vitro PTC model.

The model can be used for many applications:

• Identification of transporter-mediated renal drug clearance pathways for xenobiotics during drug development
• Identification of clinically important transporter-mediated Drug-Drug interactions during drug development and post market in clinic (drug induced AKI)
• Identification of cross species differences in renal drug handling – de-risking adverse outcomes at first in man
• Application of renal model to identify renal target and target engagement/efficacy
• Development of screening regime for biologics transport and toxicity
• Identification of drug induced kidney damage using clinically relevant biomarkers of nephrotoxicity, cross species, as a predictive tool to improve ‘first in man’ outcomes

• Proximal tubule cells are plated on 24 well Transwell plates
• Parallel assays are available using PTCs from a range of species: Human, Dog, Rat, Mouse and Non-Human Primate
• Cell monolayers undergo QC (TEER of >60 ohms.cm²)
• We measure Absorptive flux (apical to basolateral; J_ab), Secretory Flux (basolateral to apical; J_ba) and calculate Net flux (J_net= J_ba-J_ab)
• We can measure transport of either small molecules or large molecules in our system
• We can measure radiolabelled compounds using scintillation counting or non- labelled compounds by LC-MS
• We correct for paracellular flux using either radiolabelled Mannitol or unlabelled Lucifer Yellow
• We include a positive control (usually 3H-PAH or 14C-creatinine) as a QC of assay performance
• To understand mechanisms of transport or Drug-Drug interactions, we can run experiments in the presence of specific inhibitors of transport proteins.
• We provide either the raw data or a full package of transcribed and analysed data, data summary and a report

• Proximal tubule cells are plated on 96 well Transwell plates
• Parallel assays are available using PTC from a range of species: Human, Dog, Rat, Mouse and Non-Human Primate
• Cell monolayers undergo QC (TEER of >60 ohms.cm²)
• Cells can be challenged for 24, 48, 72 or 96 hours with compounds under investigation
• We measure 6 endpoints of kidney injury: KIM-1, NGAL, Clusterin, TEER, Cellular ATP content, LDH release

Our podocyte model is composed of a monolayer of freshly isolated and fully differentiated podocytes. The podocyte model recapitulates the structure and function of the glomerular filtration barrier showing size and charge selectivity which can be modulated by the addition of known nephrotoxic drugs affecting the glomerular filtration barrier such as puromycin and adriamycin.

The model is one of the first robust, fully differentiated, fresh primary podocyte model for in vitro assessment of drug-induced renal toxicity in the glomerulus.

Our unique service evaluates podocyte injury and glomerular permeability in vitro and allows our clients to assess the effect of their compounds on the glomerulus.

• Podocytes cells are plated on 96 well Transwells
• Cell monolayers undergo QC (TEER of >60 ohms.cm²)
• Cells can be challenged for 72 hours with compounds under investigation
• We measure 3 endpoints of podocyte injury: TEER, Cellular ATP content, and permeability of 70 kDa FITC-dextran

Yes, we have extensive experience of working with both small and large molecules. aProximate™ proximal tubule cells express both Megalin and Cubulin and the transferrin receptor.  We have extensive experience of measuring both transepithelial transport, cellular accumulation and potential nephrotoxicity of a wide range of large molecules including;  siRNA, Anti-sense Oligonucleotides, peptides, antibody fragments, antibiotics and other large drug molecules.

The amount of material depends on the size of the study, the molecular weight of the compound and the concentration range tested.  We can provide that information as we generate a scope of work for the project.

The turnaround time to deliver experimental results varies depending on the nature and size of the study and which species is required.  As a rough guide:

Transporter study: 3 separate biological kidneys 6-8 weeks
Nephrotoxicity Study PTC and Glomerulus : 3 separate biological kidneys 10-12 weeks

These timescales are estimates for studies involving human, rat, dog or mouse kidney.  NHP kidney is subject to availability.

In addition to transporter, DDI and nephrotoxicity assays we also offer:

• SeaHorse assay of mitochondrial health
• Generation of samples of Toxigenomic studies
• Generation of samples for RNA TempO-seq analysis
• High content screening

We use the classical fibrotic agent, TGFβ.

The primary human lung fibroblast model is only available as a service for our FMT assay. The advantage of outsourcing to us is that our experts to design the best experimental protocol.

We usually expose our cells to compounds at a range of concentrations then stimulate FMT transition for 72h and evaluate the ability of the compound to reduce ECM deposition. We recommend performing the experiments with cells from three different donors to validate the results. All  readouts are performed using High Content Imaging (HCI).

We evaluate several markers, including α-SMA (alpha smooth muscle actin A), collagen I and collagen III.