Frequently Asked Questions - Newcells

Retinal platform

An in vitro, light responsive, retinal model for accurate predictions of in vivo outcomes you can have confidence in.

aProximateTM

Kidney platform

The most advanced near-physiological high throughput kidney proximal tubule cells (PTC) model to investigate drug transport modalities in vitro.

Lung epithelia model

Lung model

A model to investigate airway physiology, viral infection, drug safety and environmental impacts on lung airway epithelia.

Sinusoid iPSC-derived Liver model

Liver model

We are developing a model of liver sinusoid derived from human induced pluripotent stem cells (iPSC).

Frequently Asked Questions

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Get answers to the most frequently asked questions.

Retina

1. Can I order ready to use products online?

For all order enquires please contact us.

2. Can you ship globally?

We are currently able to ship retinal organoids to Europe and the USA. In addition, we are performing trial shipments to Japan, please get in touch if you would like to partner with us for the trials.

3. How do you ship your organoids & RPE? 

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 currently offer RPE cells as a service and expect to ship them as a product in frozen vials in Q1 2023.

4. How do you ensure that your organoids are ‘ethically sourced’ / from ethically sourced donors?

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.

5. Can I order organoids at different ages e.g. 120, 150, 180 days?

We offer the organoids as a ready-to-use assay (RTU) at day 150 of the differentiation. Depending on customer needs, we are able to ship organoids at different ages.

6. Do retinal organoids contain RPE too?

Retinal organoids do not typically contain RPE.

7. Do you offer vector design?

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 though multiple customer and collaboration projects using our models.

8. Can you isolate the cell types in the retinal organoids and study them individually?

We are able to provide scRNAseq as a service, please contact us to discuss your specific project needs.

9. How long does it take to develop organoids with a specific disease?

Generation of CRISPR-Cas9 iPSC lines takes 12-30 weeks depending on the editing complexity required. Generation of retinal organoids takes additional 26 weeks [VC1] for day 150 organoids.

10. How long do retinal organoids last?

Retinal organoids are sold as a RTU assay at day 150 of the differentiation, however depending on the application and project requirement they can be used at later stages. Please contact us for more information.

11. Can you rescue retinal organoids post-infection?

We do not recommend using retinal organoid cultures that we compromised by infection.

12. What cell types can you reprogram into iPSCs and differentiate into retinal organoids?

We are able to reprogram dermal fibroblasts and PBMCs into iPSCs.

Kidney

1. Why use the aProximate™ in vitro proximal tubule platform?

In vitro assays  are gaining traction as New Approach Methodologies (NAMs) to reduce unnecessary animal useage in the development of new drug molecules. In vitro assays such as the aProximate™ or Microphysiology Systems (MPS) provide  near physiological models of tissues that provide  robust, reproducible  data from well qualified models  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. The aProximate™proximal tubule  platforms:

  • Are extremely well differentiated and representative of in vivo proximal tubule
  • Are supported by a large package of validation data to support their use in transporter and nephrotoxicity assays and value in predicting in vivo outcome.
  • Are available across the main preclinical species (rat, dog, NHP and mouse) together with human
  • Have been used in  Regulatory submissions to support  NDA and IND submissions
  • Are performed state of the art laboratories  by experienced scientists  working to strict SOPs  and Quality Management System

2. What are the applications of the aProximate™ proximal tubule platform?

  • 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

3. What does a typical transporter experiment look like?

  • Proximal tubule cells are plated on 24 well Transwell Filter supports
  • Parallel assays are available Human, Dog, Rat, Mouse and Non- Human Primate proximal tubule monolayer
  • Cell monolayers undergo QC before use with TEER of >60 ohms.cm2
  • We measure Absorptive flux (apical to basolateral; Jab), Secretory Flux  (basolateral to apical; Jba) and calculate Net flux (Jnet= Jba-Jab)
  • We can measure transport of either  small molecules  or  large molecules in our system
  • We can measure either  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 presence of specific inhibitors of transport proteins.
  • We provide a package of  transcribed and analysed data, summary powerpoints and a report

4. What does a typical Nephrotoxicity experiment look like?

  • Proximal tubule cells are plated on 96 well Transwell Filter supports
  • Parallel assays are available Human, Dog, Rat, Mouse and Non- Human Primate proximal tubule monolayer
  • Cell monolayers undergo QC before use with 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
  • We can measure transport of either  small molecules  or  large molecules in our system
  • We include a positive control (usually cisplatin  or polymyxin B) as a QC of assay performance
  • To understand  the role transporters or Drug -Drug interactions in modulating kidney injury response , we can run experiments in presence of specific inhibitors of transport proteins.
  • We provide a package of  transcribed and analysed data, summary PowerPoints and a report

5. Have you any experience in working with large molecules or biologics?
Yes, we have extensive experience of working with both small and large molecules. aProximate™ proximal tubule cells express both megalin and tubulin and the transferrin receptor.  We have extensive experience of measuring both transepthelial 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.

6. How much material do I need to send for a study?
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.

7. What is the timescale in delivering results?

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

Transporter study: 3 separate biological kidneys 6-8 weeks
Nephrotoxicity Study: 3 separate biological kidneys 8-10 weeks
These times are estimates for studies involving human, rat, dog or mouse kidney.  NHP kidney is subject to availability.

8. Do you offer any other assays?
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

9. What are the first steps in working with Newcells?
The first step is to contact one of our Account Managers. The Account Manager  will  be able to introduce our assays  and introduce you 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 details the study in detail and includes a statement of the deliverables form the project and a predicted timescale. 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 you point of contact throughout the study. At the end of the study the Study Manager will collate the results and the report.

10. Can we be involved in the design of the study and the report?
Absolutely!  The study design is a joint endeavour between your scientist and the Study Director and Study Manager at Newcells  to produce the best study possible. Our experience 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.

11. Can I receive updates on progress through the course of the study?
Yes, communication is very important, we can send interim results at any point in the study. Before we send results we will perform a QC of the data and results.  Equally the Study Manager is available for teleconference or Zoom meetings to discuss any ongoing study.