We derive iPSCs from peripheral blood mononuclear cells (PBMC) or primary fibroblasts (other cell types are possible, as well, upon discussion). Our iPSCs are generated using non-integrating Sendai virus carrying the “Yamanaka factors” OCT4, SOX2, KLF4, and c-MYC or mRNA transfection (fibroblasts). Extra services include isolation of PMBCs from fresh blood, isolation of fibroblasts from skin punch biopsies, Sendai clearance test, and functional validation of trilineage differentiation potential. We additionally offer technical support in rescuing difficult and unstable iPSC lines. This service includes: three iPSC clones (three vials each: 10⁶ cells/vial), a mycoplasma test report, and a panel of pluripotency markers (immunofluorescence). Please allow 6-8 weeks for iPSC derivation. Additional time is required for different starting cell types than PBMC or fibroblasts.

We offer a wide array of high-quality gene editing of iPSCs. To overcome the most common challenges of gene editing, we employ smart guides and transfection optimization (for efficient editing), transient RNP expression (to control off-targets), and smart growth and selection (for true clonal editing) in combination with continuous monitoring of morphology and pluripotency markers. We can generate knockout, knock-in (SNP insertions), large deletions, epitope tag insertions, or reporter insertions in both hetero- and homozygosity. Knockouts and knock-ins can be generated as constitutive or as conditional (e.g. Cre recombinase dependent). This service includes two iPSC clones for either KO, SNP, tag cell clones (two vials each: 10⁶ cells/vial), and an indel and knockout report (sanger sequencing). Knockouts typically take 4 to 10 weeks, while knock-ins require around 10 to 12 weeks.

The Stem Cell Engineering CoRE works in combination with the Mouse Genetics and Gene Targeting CoRE to generate transgenic mice, including knockout and knock-in by injection of gene editing reagents into mouse embryo.

To genetically characterize your iPSC, we offer an in-house ultra-fast and inexpensive report of the eight most common karyotypic abnormalities reported in human iPSC. We also provide a more in-depth karyotyping analysis based on microarrays and short tandem repeat or STR analysis for 15 loci to monitor the identity of a line.

We offer two panels to functionally characterize your iPSC: the Pluripotency Card to monitor the expression of markers associated with pluripotency, and the trilineage card to validate the ability of cells to differentiate to the three germ layers: ectoderm, mesoderm, and endoderm.

We also offer a convenient and cost-effective in-house test to detect all common mycoplasma contaminations in the supernatant of iPSC and other eukaryotic cells.

This service includes an assay report and raw data when available. Delivery Times are as follows:

• Karyotype: 1-10 weeks
• STR analysis: five-six weeks
• Pluripotency Card: two-four weeks
• Trilineage Card: three-five weeks
• Mycoplasma Test: one week

The CoRE offers differentiation of iPSC to all major lineages, including cortical and motor neurons, cardiomyocytes, hepatocytes, skeletal muscle cells, and hematopoietic lineages. This service includes two or more plates of live cultures, depending on the lineage. Depending on the protocol, please allow 3-10 weeks for delivery.

Due to the project-specific nature of each project, the CoRE offers customized services tailored to the researcher’s needs. Please use our free consultation service to inquire about your special needs.

All service rates are indicated at the point of reservation within the iLab. Please inquire for customized quotes, as editing and differentiation protocols vary depending on cells, locus, and lineage. Discount rates are offered for projects involving more than three lines.

The CoRE’s projects are carried out under a full-service modality, in which after consultation, the CoRE performs the experiment in a sufficient way. The user is only contacted at the end of the service or for major report updates.

Alternatively, we offer a host service modality in which the user is involved not only in the planning phase but also in the experimental phase. Under host service modality, personnel from the user’s laboratory and the CoRE work together to carry experiments and interpret results. Please inquire about the discount rates applicable to your host service modality.

Master PSC Lines: Master cell lines are well-known human iPSC and embryonic stem cell (ESC) lines quality-ascertained by the CoRE. They are the CoRE’s first choice for editing and differentiation protocols because of their normal and stable karyotype and their amenability to editing. Discounts are applied to projects using master cell lines provided by the CoRE.

Current master PSC lines include:

WTC-11
NCRM-2
WA09, H9
WA07, H7
WA01, H1
RUES2

MSN iPSC “Control” Lines: The choice of proper controls is critical in an iPSC experiment. The MSN lines are part of a collection of iPSC derived from healthy individuals and are available at the CoRE. All lines are authenticated by STR, characterized by IF (Nanog, Oct4, Sox2, SSEA4, TRA1-60) and Tri-lineage differentiation. The lines have normal karyotype tested by G-banding and SNP array-based karyotyping. The MSN lines are derived from fibroblasts using SeV and tested negative for detection of SeV. Clinical data, RNAseq, and Whole-genome sequence (WGS) are available. Clinical phenotypes, STR and WGS information is available at dbGAP, under restricted control-access, and RNA-seq is available from GEO. The clinical data are also available.

Additional metadata, signatures, SOPs, and other components are available on the Mount Sinai Institute for Systems Biomedicine.

Our fluorescently tagged iPSC lines are from the Allen Cell Collection. The Collection has 48 high-quality-certified fluorescently tagged hiPSC lines that target 38 key cellular structures and substructures. The WGS data (100X coverage) and RNAseq data for the parental line (WTC-11) and for all the edited lines is available.