Orders must be placed via our website. If you are a new customer, you can create an account by filling out our new customer application form. If you encounter any issues during the ordering process, please contact customer service at orderquest@jax.org or call +1.800.422.6423 (toll free from the USA, Canada, and Puerto Rico), +1.207.288.5845 (from any location). Please note that we are currently not able to ship iPSCs to China.
You can find our cancelation policy here.
As our cell lines are shipped on dry ice (within the US, Canada, Puerto Rico and Europe) or in a dry shipper containing vaporized liquid nitrogen (to any other country), additional costs apply dependent on the weight and destination of the package.
Once you have placed an order, you will receive an email with your unique case number. You will also get an email confirmation within 24 hours. Finally, you will get a FedEx number if you are in the US, Canada or Puerto Rico once your order has been shipped. For all other countries, you will be able to track your shipment via the airway bill number or via the door freight forwarder.
We ship cells on Mondays, Tuesdays, and Wednesdays. We aim to deliver your cells within two business days if you are in the US, Canada or Puerto Rico. Depending on the extent of paperwork that is required to ship cells to a country outside of the US and Canada, it may take some time until you receive your order. Our customer service team will be in touch with details following order placement.
Our cells are shipped on dry ice within the US, Canada, Puerto Rico and Europe. For all other countries, cells are shipped in a dry shipper containing vaporized liquid nitrogen. The vials need to be removed from the metal cane that sits in the dry shipper. Once received, we recommend storing cells at -80°C for temporary storage (1-5 days) and at -196°C for long-term storage.
Each vial contains 0.5 x 106 cells frozen in 0.5 ml media.
The JAX standard protocol can be downloaded from the iPSC landing page.
JAX solely functions as the distributor of iPSC lines that have been generated in collaboration with iNDI and with support from CZI. We rely on our expert partners at NIA and the scientific community to share their protocols for differentiation with us. Some have already been listed on protocols.io and have been included in Pantazis et al, 2022. While it is very difficult for us to advise on or troubleshoot specific differentiation questions, we will continue to share additional protocols and references as they become available and suggest to also review the manuscripts on bioRxiv and published papers that have used KOLF2.1J and derived lines.
The identification number on the frozen vial is different from the product code of the cell line that is displayed on the website or in your order confirmation email. To trace the vial number back to the identity of the cell line, please use the package insert that shows the vial number for each product code after Sample ID. You can use your confirmation email to associate the product code with the cell line. An example of a tube and its unique 10-digit number is in the The JAX Standard Protocol which can be downloaded from the main webpage.
You will find a FedEx return label with the shipment that should be used to return the dry shipper. To avoid additional costs, please send the dry shipper back as soon as possible.
Please note that our product codes have recently changed to include two additional zeros. Example: JIPSC1000 is now JIPSC001000.
While the product codes are changing, the products themselves, including their quality and specifications, will remain the same.
We have posted the iPSC Terms and Conditions on the webpage to allow you to review the conditions prior to requesting an order. In addition, a copy of the iPSC Terms and Conditions will be sent to you by email within one day of your order. JAX general Terms and Conditions can be found here.
Please note that we do not require you to return a signed copy of the iPSC Terms and Conditions or provide a statement on how you intend to use these lines. The iPSC Terms and Conditions is provided at the time an order is placed as a condition of purchase. Unfortunately, we cannot consider any amendments to the iPSC Terms and Conditions as it was drafted with the input and consent of multiple parties with rights or interest in the iPSC lines/related IP.
If cell viability is less than 80% or you encounter any contamination within 24 hours of cell culture for the pathogens tested in the Certificate of Analysis, please contact our Customer Service team immediately at orderquest@jax.org or call +1.800.422.6423 (toll free from the USA, Canada, and Puerto Rico), +1.207.288.5845 (from any location).
If you have any technical questions, please email our technical information scientists at micetech@jax.org.
The DNA sequence in the parental cell line is considered the reference and the wild type (WT) allele designation reflects this. The gene-edited variant allele is noted as SNV (single nucleotide variant). The revertant allele (Rev or REV) is the same sequence as the parental allele (WT) and was derived from the gene editing process of modifying the SNV allele back to wild type.
Each cell line product page contains downloadable Sequence Map files. The exact DNA change is annotated in the Features sections of these design files. The information is readily found in the SNV subsection of the Features section which specifies the nucleotide base and its relationship to WT, REV or SNV cell line name nomenclature.
Most mutations in this catalog are associated with single nucleotide variants defined by the NIH National Center for Biotechnology dbSNP’ database. This unique identifier is present in the Genomic Features details of each cell line’s details page. The exact genomic coordinate of the variant is indicated in the Genomic Features section as well. Finally, the Sequence Map files for each cell line contain rich data and visual presentations of the gene features, gene editing tools, and genotyping primers, as well as DNA/Protein information.
The Sequence Map for each cell line is created using SnapGene software by importing the current human reference genomic sequence gene (GRCh38), RNA and protein annotations. The cell line-specific gene editing reagents, genotyping primer sequences, Sanger Sequencing primer(s), and variant specifics are included and presented in a comprehensive visual map. These Map files are available in three different file types: SnapGene, PDF and GenBank. NOTE: The reference sequence in these Maps does NOT represent the sequence of the KOLF2.1J parental genome sequence.
You can find more detailed information in the Sequence Map document. The SnapGene® software package has been used to create the Sequence Map files (snapgene.com). A free read-only version of the software can be downloaded for viewing the SnapGene file type. In the PDF output of the SnapGene file type map, the first page displays a general overview of the respective gene derived from the human reference genome database (gray). The canonical primary Ensembl RNA transcript used in this Map is shown in orange, the protein coding sequence is displayed in purple, the donor template is shown in sky blue, the protospacer in red, and the SNV in orange. The subsequent pages show the detailed sequence in base pair resolution with the respective features annotated including the sequence of the PCR primer pair used for Sanger sequencing validation. The base in the reference sequence targeted for change is colored in red letters.
Genes and variants were selected by the researchers composing the NIH iNDI. The Ramos et al, 2021 publication in Neuron provides a more detailed context.
We will be adding additional cell lines to our existing Distribution Sets once available, including whole gene knockouts and SNV-containing and wild type genes fused to the multifunctional HaloTag sequence. The aim is to generate cell lines for 134 variants across 73 ADRD genes. Please email micetech@jax.org if you are looking for a variant that is currently not covered.
You can find this information on our website catalog by clicking on 'view more' under ‘gene editing sequences’ of the respective cell line. The protospacer sequence of the gRNAs is also included in the Sequence Map files.
You can find this information on our website catalog by clicking on ‘view more’ under ‘DNA Primers for Sequence Validation’ of the respective cell line. The primers used for PCR and Sanger sequence validation are also included in the Sequence Map files.
The use of revertant lines may allow discrimination of phenotypes caused by the mutation to not be confused by off-target consequences of the gene editing process to create SNVs. Undetected off-target mutations are unlikely to be correctly gene engineered back to the KOLF2.1J “reference” state (Wildtype=WT). If the phenotypic signal is not suppressed in the revertant, it is possible the experimental signal detected is not caused by the expected SNV but by a second mutation.
In the ‘Genetics’ section of each cell line’s details page, the genotype of the revertant’s parent is noted in the ‘Origin of Revertant’. For much of the catalog, the autosomal dominant mutations have reverted heterozygous lines, while for recessive mutations, we have engineered homozygous lines back to the wildtype (KOLF2.1J) sequence. Some JAX Distribution groups will have revertants created from both heterozygous (SNV/WT) and homozygous (SNV/SNV) cell lines.
During the cell line creation process, gene edited clones containing the desired DNA sequence were subjected to further quality control studies. Karyotype (G-Banding) analysis was performed and only clones with high percentage euploidy and normal G-banding patterns were allowed to proceed to the production scale stage, and are provided in this catalog. The report can be downloaded from each cell line’s webpage and can be found in the QUALITY CONTROL DATA section.
Whole-genome sequencing has NOT been performed. However, genome-wide SNV array technology has been used to interrogate the copy number variation of the entire genome landscape. Analysis of genomic DNA on Infinium Global Diversity Arrays (GDA) with added NeuroBooster content was performed as previously described [1]. In brief, GDA provides superior coverage across the genome with ~two million variants and with the added NeuroBooster content, which includes the majority of known neurodegenerative disease SNPs. DNA was isolated from iPS cell lines and used for genotyping using the GDA+NeuroBooster array with standard Illumina genotyping protocols. To assess genomic integrity of iPS cell lines, we investigated the B-allele frequency and Log R ratio values of each iPS cell line, which were downloaded from Illumina GenomeStudio. Abnormal patterns were observed by visual inspection and were plotted using R (v3.6.1) with the GWASTools package [2]. Additionally, genotyped cell lines were compared to the KOLF2.1J WGS data and investigated for large patterns of mismatching SNPs which implied a genomic integrity issue. Scripts and code can be found on the Center for Alzheimer's and Related Dementias (CARD) iNDI project Github.
Only cell lines post gene-engineering process that passed the DNA sequence, Karyotype, and SNV Array procedures were expanded for distribution. The cells expanded and provided to customers have had additional quality control performed on them. The distribution LOT quality control procedures are as follows:
The parental line KOLF2.1J and its mutant derivatives carry heterozygous small copy number variants (CNVs), which have been described in detail by Gracia-Diaz et al.
While we acknowledge the presence of these CNVs in KOLF2 lines, to date, we have not identified any impact on pluripotency nor differentiation capabilities. KOLF2.1J and mutated lines were shipped to over 100 labs prior to release, with the aim of testing KOLF lines in various functional assays and probing performance against several in-house reference lines. Researchers collectively found that KOLF2.1J was able to support a variety of neuronal differentiation protocols at efficiencies similar to other well-known iPSC lines, arguing that these specific and small CNVs are neutral for neuronal development (https://www.cell.com/cell-stem-cell/fulltext/S1934-5909(22)00451-9).
Additionally, the variants chosen for the iNDI project are generally fully penetrant genetic variants (i.e., the variants are likely to be causal for disease irrespective of genetic background). Therefore, we currently have no concerns using KOLF2.1J and derivatives for research into highly penetrant variants which are causal for age-associated neurodegenerative conditions. We encourage end users to evaluate if their specific experiments are likely to be impacted by these CNVs, in particular potential interaction with other variants researchers may wish to introduce.
For more details, please refer to the letter we recently published in Cell Stem Cell: 'Large structural variants in KOLF2.1J are unlikely to compromise neurological disease modeling.'
The distribution LOT of cells customers receive have NOT been evaluated in pluripotency protocols. However, the KOLF2.1J parental cells were evaluated at a similar passage number to that used to initiate the gene editing procedure. Furthermore, early access customers have used a limited number of the gene edited cell lines in their own research facilities and have found them to be pluripotent and functional in their differentiation protocols. You may find more information here: https://www.cell.com/cell-stem-cell/fulltext/S1934-5909(22)00451-9
The Certificate of Analysis, which can be downloaded from each iPSC line webpage, indicates the range 'p4 through p6'. The exact passage number of the line you are receiving will be added to the package slip that comes with the vials.
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