Who are NINDS?

The National Institute of Neurological Disorders and Stroke (NINDS) was established in 1950 and is a part of the US National Institutes of Health (NIH). Its mission is to seek fundamental knowledge about the brain and nervous system to reduce the burden of neurological disease for all people.

NINDS aims to provide tools and funding for research into neurological disorders that cause a considerable burden on individuals, families, and communities. NINDS research funding has already led to the development of precise diagnostic tools and new treatment interventions for many neurological conditions. The institute also invests heavily in educating the general public and medical practitioners on the signs and symptoms of neurological disorders and stroke.

One of NINDS’ critical projects is the extensive collection and storage of human cells that can be requested for research purposes. This extensive collection of cells come from a large population of donors including men, women and children of all wide and diverse range of backgrounds. The repository of biomaterials is not restricted to researchers based in the US and can be requested by small academic groups through to large biotechnology and pharmaceutical companies.

NINDS Human Cell and Data Repository (NHCDR)

The NHCDR hosts over 400 subjects that can be requested for research purposes. These subjects are sourced from a wide array of hosts and can be filtered for by gender, age, race and cell type (iPSCs, edited iPSCs or Fibroblasts). Subjects can also be selected based on the disease type, of which there are 14 to choose from including:

• Alzheimer’s disease
• Amyotrophic Lateral Sclerosis
• Ataxia-Telangiectasia
• Dystonia
• Frontotemporal Degeneration
• Huntington’s Disease
• Lewy Body Dementia
• Mild cognitive impairment
• Myotonic Dystrophy
• Parkinson’s Disease
• Parkonsonism
• Spinal Muscular Atrophy
• Spinal-Bulbar Muscular Atrophy
• Spinocerebellar Ataxia
• Control samples

A full comprehensive list of subjects held by NINDS can be found on their website. If you wish to query or request cell samples, an easy “how to order” guide can be found here. Once a Material Transfer Agreement form is completed (found on NINDS website here), researchers can search for the cells they require and purchase up to a maximum of 10 vials.

NINDS impact on research

The NHCDR has supported a diverse array of stem cells-based research, from studies of the basic biology of stem cells in the developing and adult mammalian brain, to studies focusing on nervous system disorders such as ALS or spinal cord injury. Other examples of research include using induced pluripotent stem cells (iPSCs) to derive dopamine-producing neurons that might alleviate symptoms in patients with Parkinson’s disease and using embryonic stem cells to generate cerebral organoids to model Zika virus infection.

Since the NINDS repository was established, countless papers have been published that have credited the organization for providing the biomaterial in their investigations. Many of the papers crediting NINDS for providing the biomaterial utilized in their investigations include high impact factor journals such as

• Nature
• Cell Stem Cell
• Molecular Cell
• Neuron
• Science Translational Medicine

Requesting cells from Sampled

Sampled is proud to be the sole distributor for the NHCDR. Although NINDS is based in the US, the biomaterials can be requested by scientists worldwide by small academic groups to large pharmaceutical organizations. Requesting cells from the NHCDR is a straightforward process, researchers can contact us directly through our website here, to speak to one of experts. We are here to help researchers, which is why we also have a step by step guide to ordering cells from the NHCDR.

The NINDS biobank stem cell collection currently contains 411 subjects, 169 of which have iPSC and 270 of which have fibroblasts and represents several neurodegenerative disorders, including (but not limited to):

• Alzheimer’s disease
• Amyotrophic Lateral Sclerosis
• Frontotemporal Degeneration
• Huntington’s disease
• Parkinson’s disease

If you would like to browse and order cell lines, first go to: https://stemcells.nindsgenetics.org/ and follow the instructions below:

Creating a signed Material Transfer Agreement (MTA)

A key requirement for ordering lines is ensuring a signed MTA is in place.

The MTA forms can be found in the Documents section of the catalog application as shown below.

Selecting cell lines

1. Once you have downloaded the correct MTA form, select the Subjects tab and choose subjects of interest using the filters to narrow by disease, etc, if necessary

Locate the desired cell lines in the subject’s detail view, clicking anywhere within the row will bring up the cell line’s details.

If the subject has cell lines, such as iPSCs and Fibroblasts, they will be displayed in the subject’s detail view. These are found at the bottom of the Full Record tab, and also the Cell Lines tab. Click the Add to Cart button for the desired lines.

Purchasing cell lines

2. Once you have selected the cell line, scroll down and Click Add to Cart.

3. You can then click on the up and down arrows to specify how many vials you would like to purchase (the maximum is 10):

4. Go to the cart and click Checkout.

5. Enter the required fields and upload the required documents, such as signed MTAs (found in the Documents section) and Statement of research intent (SRI).

6. Once all relevant information has been filled in, Click Submit Order 

7. You will receive an e-mail summarizing your order and requesting e-mail address verification.

Here is an example of the e-mail you will receive. Click the link to verify your email address and your order should be all set.

Cell and Data Repository infographic

Download our infographic

View further details on our catalog of the NINDS Human Cell and Data Repository, including a breakdown of gender, race and clinical affected status of cell and disease types.

View infographic

If you need any help ordering from the NINDS website, you can contact us at NINDS@sampled.com and one of our experts will be happy to help.

You can also view our other cellular services here.

How does CRISPR gene editing work?

When CRISPR Cas9 protein is added to a cell, along with a strand of guide RNA, the Cas9 protein first binds to the guide RNA. This complex is then able to move along the DNA sequence until it locates a specific sequence of DNA (approximately 20 base pairs long) that matches the guide RNA. Once the complex locates the specific sequence or gene it was programmed to, it cuts the DNA before mutations are introduced to make precise changes to the DNA sequence. This can include removing the sequence or turning the genes on/off by changing the one letter of the DNA code to another.  

The promise of CRISPR technology is very exciting as the technology opens doors for groundbreaking changes to everyday life, including the prevention and treatment of many diseases, increasing crop yield and even editing out harmful genes within the DNA of fetuses.

So, what are the benefits of outsourcing CRISPR gene editing?

While some research institutions have developed in-house expertise in CRISPR gene editing, there are several compelling reasons why outsourcing these services can be beneficial. In this blog, we will explore some of the five reasons why outsourcing CRISPR gene editing services can be advantageous.

Access to specialized expertise

CRISPR gene editing is a complex and rapidly evolving technology that requires specialized expertise to execute with precision and accuracy. With CRISPR being a new technology, there is a lack of user friendly guides or resources to help researchers who wish to carry out CRISPR by themselves. This has led to scientists who are unfamiliar with protocols attempting their experiments several times before achieving some success, with many citing that the optimization step is particularly difficult to execute properly. The success achieved by the few who have managed to delete/insert genes is limited however, as editing efficiency is far lower than they would hope.

By repeating the same experiment several times, only to obtain low editing efficiency, researchers who have tried to execute CRISPR research are wasting not only time and resources but money too. Outsourcing gene editing services to a specialized service provider such as Sampled allows researchers to gain access to a team of experienced and knowledgeable experts with deep expertise in this field. We  are equipped with the latest gene editing tools and technologies, allowing us to deliver efficient and effective solutions for a wide range of gene editing applications.

Cost-effective solution

Outsourcing CRISPR gene editing services can be a cost-effective solution for research institutions and biotech companies. The cost of setting up an in-house gene editing laboratory can be high, as it requires significant investments in equipment, personnel, and training which can take a long time. By outsourcing gene editing services, institutions and companies can save on these costs and leverage the expertise of specialized service providers who have already made these investments.

Flexibility and scalability

Outsourcing CRISPR gene editing services to Sampled offers flexibility and scalability. We can quickly adapt to your project needs and timelines changing, providing researchers with the ability to scale up or down their gene editing projects as required. This flexibility can be particularly valuable in situations where resources are limited or where project timelines are subject to change.

Quality assurance

Outsourcing gene editing services can also provide researchers with quality assurance and peace of mind. Our tried and tested established protocols and quality control processes are in place to ensure that all gene editing projects are executed to the highest standards of accuracy, efficiency, and safety. By outsourcing gene editing services, researchers can be confident that their gene editing projects will be executed to the highest standards, providing reliable and reproducible results.

Regulatory compliance

CRISPR gene editing is subject to stringent regulatory compliance requirements, particularly when it comes to clinical trial research. By outsourcing gene editing services to a service provider that specializes in regulatory compliance, researchers can ensure that their projects meet all regulatory requirements and standards which in turn ensures your project won’t suffer delays. This can be particularly important for researchers seeking to develop new gene therapies or other clinical applications of CRISPR gene editing.

The benefits of outsourcing CRISPR gene editing work to Sampled

We accept induced pluripotent stem cells (iPSCs) directly from clients or those selected from repositories housed at Sampled such as National Institute of Mental Health (NIMH; https://www.nimhgenetics.org/) and the National Institute of Neurological Disorders and Stroke (NINDS; https://stemcells.nindsgenetics.org/). Our trained experts can discuss your needs and help with experimental design so that you can be assured that the cells to be edited meet your expectations. Furthermore, our cells undergo rigorous quality control testing to ensure that you will receive a 100% homogeneous population, this is crucial for investigations such as drug screening studies to minimize noise and prevent genetic drift within cell populations.

To summarize, outsourcing CRISPR gene editing services can provide many benefits to research institutions and biotech companies. By gaining access to specialized expertise, cost-effective solutions, flexibility, quality assurance, and regulatory compliance, researchers can execute their gene editing projects with confidence and achieve their desired outcomes, without having to become CRISPR experts overnight.

To get in touch with our experts and learn more about our Sampled Labs Services click here.

Induced pluripotent stem cells (iPSCs) derived from human somatic cells hold massive potential for research into disease-modelling, drug toxicity testing and in regenerative medicine. Thus, as the use of patient-derived stem cells has become more frequently adopted, having a workflow to monitor each cell line is critical. For these reasons, it is imperative that they pass a series of quality control measures to promote reproducibility across experiments and between labs.

To consider these needs, a multistep workflow to assess newly generated iPSCs is vital to test four critical benchmarks:

1. Viability and sterility of cells
2. Genetic abnormalities
3. Stemness
4. Pluripotency (ability to differentiate into the three germ layers)

Testing the sterility

Before using any type of cell line for research it is necessary to ensure that the cells in question are viable and are not contaminated. The biggest entry of species such as Mycoplasma into cell lines are from non-sterilizable cell culture reagents and laboratory personnel. M. orale, M. fermentans, and M. hominis account for more than half of all mycoplasma infections in cell cultures and are found in oropharyngeal tracts¹. Two other species that are known to be prevalent in laboratories are M. arginini and A. laidlawii which can be found in Fetal Bovine Serum and swine-derived Trypsin solutions. Unfortunately, mycoplasma implies resistance against penicillin and can pass 0.2μm sterility filters². This makes it extremely difficult to eradicate mycoplasma infections, furthermore numerous research laboratories do not test for it on a regular basis.  At Sampled however, all iPSC lines are tested for mycoplasma.

G-banded karyotyping and testing for genetic abnormalities

Once the cells have been tested for sterility, it is then necessary to test for genetic abnormalities that could have been introduced into the cells’ DNA during the reprogramming process or induced through culture, giving certain cells a growth advantage. Advances in the field have led to “foot-print-free” reprogramming methods that do not result in reprogramming factor integration, but these new methods do not preclude culture induced genetic mutations. Regardless of which method your iPSCs have been derived, if the cells’ DNA has been compromised, the cells may not be fully functional or perform inadequately.

Furthermore, long-term iPSCs culture can result in chromosomal abnormalities, modification of gene expression, as well as increases the risk of the iPSCs becoming tumorigenic. Since genomic alterations present potential risks in downstream applications, it is important to monitor the genomic integrity of these iPSCs lines especially in iPSCs intended for therapeutic use. It is for these reasons that iPSC karyotype analysis is necessary not just at the start, but throughout your investigation.

Measuring stemness

Immunostaining alone is inefficient at determining stemness since it is difficult to quantify immunostaining results over an entire culture.  At Sampled, we quantitate a large range of stemness markers using Pluritest. This assay uses a the Affymetrix PrimeView Global Gene Expression Array to confirm the undifferentiated state of human pluripotent stem cells.

Fluorescence activated cell sorting analysis (FACS) is another useful method that is commonly employed to measure stemness by measuring expression of stemness markers quantitatively over a large population. FACS relies on the binding of highly specific antibodies tagged with fluorescent dyes to cells expressing stemness markers such as Oct4, Tra-1-60, etc.

Measuring differentiation potential

Lastly, the iPSC’s ability to differentiate into the three germ layers (Ectoderm, Endoderm and Mesoderm layers) ought to be measured before they can be utilized by investigations. In order to test this, the cells need to first be differentiated into Embryoid bodies (EBs). Once the cells have undergone differentiation, the cells are then harvested for gene expression analysis by qPCR and analyzed to ensure that genes specific to all 3 germ layers are expressed.

Is there an easier way to validate your Induced Pluripotent Stem Cells?

It may be daunting for a small or scaling biotech to conduct the tests outlined above before they even begin to start working with these cells. At Sampled, we offer robust quality control services, that can be bundled as part of a package or as bespoke services that includes:

• Sterility testing of all lines includes mycoplasma testing
• Identity verification using the SNPTrace Panel
• Analysis of stemness by FACS
• Demonstration of stemness by Pluritest
• Demonstration of differentiation potential by hPSC scorecard
• Assessment of genomic stability by G-banded karyotyping
• Demonstration of loss of Sendai virus after reprogramming by qPCR

Our highly trained scientists perform these tests to the highest standard and could save your lab time and resources so that you can focus on the research that matters to you the most. If you would like to speak to one of our experts today about validating your stem cells, contact us today.

References

1. Pamies, D., Bal-Price, A., Simeonov, A., Tagle, D., Allen, D., Gerhold, D., Yin, D., Pistollato, F., Inutsuka, T., Sullivan, K. and Stacey, G., 2017. Good cell culture practice for stem cells and stem-cell-derived models. Alternatives to Animal Experimentation: ALTEX, 34(1), pp.95-132.
2. Bruchmüller, I., Pirkl, E., Herrmann, R., Stoermer, M., Eichler, H., Klüter, H. and Bugert, P., 2006. Introduction of a validation concept for a PCR-based Mycoplasma detection assay. Cytotherapy, 8(1), pp.62-69.