Our scientists have extensive experience using patient-specific PBMCs or fibroblasts to generate induced pluripotent stem cell (iPSC) lines. We have perfected both footprint-free (episomal, mRNA and other methods) and retroviral reprogramming methods for iPSC generation.
The advantages of using our footprint-free Custom iPSC Generation Service:
Highly efficient reprogramming (0.2%-0.5%)
High quality and purity iPSCs
Footprint-free reprogramming of patient-derived samples: PBMCs (blood)/ fibroblast
Well-optimized feeder/ feeder-free protocols
Suitable for drug discovery and cell therapy applications
ASC is one of the first, licensed service providers of iPSC and CRISPR/Cas9 technologies, and provides the best iPSC disease modeling service on the market.
- >98% success rate in genetically engineering >500 unique iPS cell line models
- iPSC genome editing using healthy/ diseased donors
- Customizable deliverables: Heterozygous/ homozygous mutations; Point mutations with/ without silent mutations
- Footprint-free, feeder-free transfection and iPSC culture protocols
- Use your iPSC lines or any of our fully characterized control iPSC lines derived from cord blood (male: ASE-9109; female: ASE-9110) or female fibroblasts (ASE-9209) for CRISPR gene editing
- Isogenic control lines for reliable experiments
- Full range of downstream iPSC differentiation and characterization services
Stringent molecular and functional assays are necessary to evaluate pluripotency and to rule-out genetic aberrations due to reprogramming and stress from in vitro culture in iPSC lines. Loss of genetic integrity can affect desired cellular phenotype and compromise interpretation and translation of results to a clinical setting.
ASC offers comprehensive services needed to completely characterize your human and mouse pluripotent stem cell lines (PSCs):
- Pluripotency and lineage-specific marker immunostaining
- Karyotyping (Chromosome counting, G-banding, array analysis)
- qPCR, RNA-seq
- Trilineage differentiation potential: embryoid body (EB) formation, qRT-PCR
- HLA typing, whole genome sequencing (WGS), single tandem repeat (STR) genotyping, copy number variations (CNV) and more
Custom “Master” iPSC Cell Line Generation using TARGATT™ technology! The TARGATT™ Master Human iPSC Line contains an attP “docking site” at the hROSA26 safe harbor genomic locus. Any gene of interest can be stably inserted at the docking site using a corresponding “attB” containing TARGATT™ donor plasmid, with guaranteed gene expression. The efficiency of this phiC31 integrase-mediated recombination is up to 100% efficiency with drug selection and up to 30% without drug selection.
Our cell line model generation experts can also engineer your safe harbor locus knock-in iPSC lines using CRISPR/Cas9 for a multi-technology approach to generate a cell line model for your specific research needs.
The true potential of iPSCs and ESCs rest in its directed differentiation to different somatic lineages. As part of our comprehensive stem cell service platform, researchers can leverage our expertise in stem cell technologies and our cost-effective, reliable Stem Cell Differentiation Services for their developmental biology, disease mechanisms, and drug discovery research.
Microglia and Neural Stem Cell Differentiation Categories
- Differentiate your patient-derived iPSCs into microglia and self-renewing, multipotent neural stem cells (NSCs) and further to neurons and glial cells.
- High differentiation efficiency and cell purity
- Cells fully characterized for microglia and NSC biomarkers
Applied StemCell, Inc. provides a comprehensive range of service to differentiate your iPSCs into cardiomyocytes using our proprietary induction protocol and reagents. We deliver ready to use, highly pure (>90%), functional cardiomyocytes. These differentiated cells provide a highly desirable in vitro platform for high content toxicity and drug screening and as a feasible alternative to animal and embryonic stem cell models.
We can generate differentiated cardiomyocytes from your own pluripotent cell lines derived from healthy or disease iPSCs.
Applied StemCell provides custom differentiation of your induced pluripotent stem cells (iPSCs) into high-quality hepatocytes for developing liver disease models, discovering therapeutic treatments, drug targets, and understanding drug induced hepatotoxicity.
Using our proprietary induction protocol and reagents, we can generate differentiated hepatocytes very efficiently and with high purity from your own healthy or disease iPSCs.
Our Hepatocytes Differentiation Service includes:
- Expansion of a host-derived iPSCs
- Progenitor cell differentiation
- Characterization of progenitor cells
- Terminal differentiation
- Characterization of Progenitor Cells/ Differentiated Cells by ICC (per marker)
Human iPSC-derived retinal pigment epithelium (RPE) provides a physiologically relevant cell line model to understand basic ocular biology and ocular diseases. ASC’s iPSC-derived RPE protocol provides high quality RPEs with expression of lineage-committed markers and functional phenotype such as phagocytosis, and by providing an unlimited source of these cells.
- RPE-like cells with typical cobblestone morphology and pigmentation
- High quality and purity cells (60-80% purity) expressing RPE-specific markers Bestrophin 1 (BEST1)/ RPE65
- iPSC reprogrammed from fibroblast/ PBMC/ CD34+ cord blood cells
- From healthy/ disease/ engineered iPSCs
- Optional! Control lines differentiated from “master” iPSC lines available
Applied StemCell has a staff of talented scientists that can help you with the more complex to the daily laboratory tasks required of iPSC cell culture. Give your staff the freedom to focus on other project, while Applied StemCell takes care of your cell culture needs.
We offer standard and customized cell culture services including mycoplasma testing and FBS lot evaluation.
- iPSC expansion services
- iPSC scale-up services
- Neural Stem Cells, Differentiated Cells scale-up services
- 3D-culture and morphology testing
Please contact us to discuss your needs with our technical service specialists.
We Understand Cell Lines! Leverage our expert, comprehensive CRISPR/Cas9 cell line modeling service for a stress-free research. With 1300+ unique cell line models engineered from >200 distinct mammalian cell lines, we can engineer perfectly suited cell line models for your research using highly optimized and efficient CRISPR/Cas9 genome editing strategies and protocols:
Cell Line Model Generation Categories
- Engineer a variety of mutations; or correct mutations, including fusion gene(s)
- Variety of cell lines: cancer, hard-to-transfect, blood lineage, stem cell lines and many more
- Start-to finish cell line generation workflow; optional downstream cell line validation
- Custom deliverables: homozygous/ heterozygous clones; point mutation with/without silent mutation
- >97% success rate; turnaround time as fast as 2 months
Cell Line Model
CRISPR/Cas9 Cell Line
Service - Hematopoietic Cells
(Jurkat and TF-1)
CRISPR Gene Fusion
Cell Line Generation
ASC is one of the earliest service providers of CRISPR/Cas9 technology, and has successfully delivered >500 CRISPR mouse models in as little as 3 months. Our animal model portfolio offers competitive pricing and turnaround times for generating CRISPR knockout, conditional knockout, locus-specific/ safe harbor knock-in, controlled gene expression and gene correction, and more.
Mouse Models Categories
- Most up-to-date CRISPR designing strategies and protocols
- 100% target-site cutting efficiency using optimized, proprietary gRNA validation methods
- AAALAC-accredited animal facility
- Animal IP belongs to researchers
- Project management and scientific support to discuss your project needs and suitable strategic options to fit your budget
- NEW! Downstream electrophysiology and behavioral assessments for your mouse models
CRISPR Conditional Knockout
CRISPR Knockout, Knock-in,
Point Mutation Mouse Models
Homologous Recombination Conditional Knockout Mouse Model and Knock-in Mouse Models
Transgenic Mice Models
With the need to physiologically relevant animals models on the rise for more predictive research findings, genetically engineered rat models are becoming increasingly popular. Get ahead of your competition by leveraging Applied StemCell’s expertise in CRISPR/Cas9 and TARGATT™ genome editing technologies and animal models engineering to generate the ideal rat model to meet your specifications and budget.
Rat Models Categories
- Global leader in genetically engineered rat models (Nature article, “CRISPR mouse model boom, rat model renaissance”)
- Rat models are generated in the US in an AAALAC accredited animal facility using ISO:9001 quality standards
- Detailed discussion to understand project requirements, optimal project design for precise genetic modification and dedicated project management for timely updates
CRISPR Knock-In, Knock-Out,
Conditional Knock-Out Rat
Bacterial Artificial Chromosome (BAC) Knock-in and Conditional Knock-In Rat Models
Applied StemCell’s proprietary TARGATT™ knock-in mouse technology enables highly efficient and site-specific gene integration to generate transgenic mouse models. This technology uses PhiC31 integrase to insert any gene of interest into a specific docking site that has been pre-engineered into an intergenic and transcriptionally active genomic locus for guaranteed transgene expression.
Advantages of TARGATT™ Technology for Knock-in Mouse Model Generation:
- Safe harbor locus “large fragment knock-in” (up to 22 kb)
- High efficiency insertion (up to 40%)
- Single copy in an active locus: avoid gene silencing and genomic instability.
- Germline transmitted F1 mice in 5-8 months
- Tissue-specific/ ubiquitous, controlled/ inducible expression options
Please contact us to discuss your project plan.
Applied StemCell’s proprietary TARGATT™ technology enables generation of physiologically relevant transgenic rat models suitable for a variety of applications including reporter gene expression, gene knock-down, conditional gene expression and disease models. This technology uses the Phic31 integrase to mediate an irreversible integration of large transgene(s) into a preselected, safe harbor locus with guaranteed gene expression.
- Site-specific, single copy transgene integration overcomes challenges associated with random integration
- TARGATT™ knock-in rats in 6-9 months
- Direct microinjection of the TARGATT™ reagents into rat zygotes
Using the TARGATT™ technology, ASC has also developed Neural Specific Cre-Rat Lines in a Sprague Dawley rat background. Please inquire.
Applied StemCell’s proprietary site-specific TARGATT™ technology can be used to generate stable, knock-in of large transgenes in cell lines, including stem cells, very efficiently and quickly. Knock-in is mediated by PhiC31 integrase at a pre-engineered “docking site” in an intergenic, transcriptionally active genomic locus (safe harbor locus) for guaranteed gene expression without disruption of internal genes. This technology allows only a single-copy integration with very high efficiency with or without clonal selection.
Use our TARGATT™ technology to generate your ”Master” cell lines, reporter/tag lines, for iPSC generation, conditional gene expression models and more.
Also, try our ready-to-use TARGATT™ Master Cell Lines to knock-in transgenes in your own lab.
The TARGATT™ technology enables faster and efficient site-specific integration of large DNA fragments in cell lines. This technology offers an ideal platform for generating stable isogenic cell line libraries for mammalian cell display-mediated antibody engineering, protein evolution screening, mammalian two-hybrid (M2H) screens and more. It allows only a 1:1 variant-to-cell ratio, and guarantees uniform and consistent expression of the gene/protein for efficient screening.
- Single copy knock-in: 1 cell, 1 docking site, 1 inserted transgene
- Site-specific knock-in: avoids problems associated with random insertion
- Isogenic cell library: comparison of phenotype against identical baseline
- Cost-effective and efficient library construction: no virus packaging time and resources
- BSL1 compatible
TARGATT™ Chinese Hamster Ovary (CHO) Cells for Recombinant Proteins and Antibody Bioproduction
Applied StemCell has created Master TARGATT™ CHO cell lines for the rapid creation of new lines for high level protein and antibody expression. The TARGATT™ CHO master cell lines yield >2.5g/L of recombinant proteins in a 2-week fed-batch shake flask expression system and is easily scalable for large scale bioproduction.
Our Master TARGATT™ CHO cell lines exceed the capabilities of the traditionally made CHO antibody production cells and also reduces the cost of bioproduction and manufacturing. Our technology makes CHO antibody production feasible and economical for companies and projects of all sizes.
Please let us know how we can help you with your CHO bioproduction project!
TARGATT™ Rabbit for Biopharming and High-Yield Recombinant Protein and Antibody Production
Animal bioproduction uses transgenic animal mammary gland as a bioreactor for the production of recombinant proteins. Animal bioproduction, compared to CHO bioproduction, has the advantages of lower upfront and maintenance costs; is easy to contain, control and transport; involves faster development processes and no scale-up issues; and has a unique low-cost bulk holding stage (frozen milk). However existing technological methods such as random integration to create transgenic animals have many limitations. Applied StemCell has overcome these limitations with our proprietary TARGATT™ technology to reproducibly, and consistently, control the knock-in locus, copy number and expression of the target protein in transgenic animals.
Applied StemCell provides lentiviral and retroviral custom virus packaging services for your CRISPR/Cas9 components, CAR-T expression vectors and other transgenes, for efficient delivery into cell lines (such as hard-to-transfect cell lines, primary cell lines) as well as direct injection into animal models.
- Rapid, custom lentivirus or retrovirus virus packaging
- Ready-to-transduce viral particles provided to you
- High quality, high titer viral particles in 10 days
- VSV-G pseudotyped viruses that exhibit broad tropism across a range of cell types
- High titer amplification of viruses, up to 10^9 infectious units per ml
- Accurate viral titers quantified by qRT-PCR
- Confirmation by DNA sequencing
- Large scale production, clonal expansion and cryopreservation
Applied StemCell is one of the earliest licensees and provider of CRISPR/Cas9 genome editing services. We are experts in designing and cloning components for CRISPR-based gene editing in cell lines and animal models using benchmark strategies and optimized protocols. Our scientists have designed > 1000 CRISPR projects for researchers worldwide, and we use the latest sequencing and next generation sequencing (NGS) protocols to validate your gRNAs and to ensure high efficiency gene modification in your gene of interest.
We offer CRISPR/Cas9 vector design, construction and gRNA validation for:
- Cell line gene modification
- Microinjection into embryos for animal model generation
- Virus packaging
We also offer comprehensive custom CRISPR cell line and mouse/ rat model generation services.
Applied StemCell has extensive expertise in anything DNA-related. We can develop the best strategy to tackle your cloning projects, and find solutions to all your technical problems.
- Comprehensive services for genome engineering for in vivo and in vitro applications
- Restriction fragment cloning
- Design and validation of CRISPR/Cas9 components: gRNA design and in vitro validation by next generation sequencing (NGS)
- Gene targeting vectors for homologous recombination
- Design and construction of vectors for transient mammalian transgene expression
- Bacterial artificial chromosome recombineering for large fragment insertion
- Design of RNAi and inducible vectors, gene tagging, and site-directed mutagenesis
- Vector design for generation of random transgenic cell line and animal models
Bacterial Artificial Chromosomes (BACs) are ultra-low copy vectors that can hold up to 300 kb of genomic fragments, making them ideal vectors for introduction of entire genes including the regulatory regions for disease modeling with transgenic animals. BACs are traditionally difficult to modify with restriction enzymes and ligases because of their large size. Applied StemCell uses homologous recombination to introduce precise changes into BACs for your specific experimental needs. Virtually any desired modification can be introduced into a BAC, including insertion of reporter genes, point mutations, Lox-STOP-Lox conditional modifications, and more.
Our custom BAC services include:
Applied StemCell can also help you create transgenic rats using BACs.
Make Informed Go-No-Go Decisions Early in Your Drug Development Process!
Leverage Applied StemCell’s unique expertise in CRISPR/Cas9 and induced pluripotent stem cell (iPSC) technologies to engineer predictive and reliable in vitro disease models as an alternative to animal models for preliminary drug screening.
Our ISO:9001 certified service platform offers full flexibility in choosing assay modules with a wide-range of functional endpoints for early-stage in vitro screening of preclinical drug candidates. We have a comprehensive cell-based test battery from which you can choose assays for efficacy, safety or target discovery that suit your therapeutic pipeline. We provide the scientific expertise, accurate and efficient screening to help you make informed decisions about your small/ large molecules early in your drug development process.
As a long-standing leader in genetically engineered animal models, Applied StemCell also offers a comprehensive custom research solutions platform for downstream assays in mouse and rat models. Our state-of-the-art vivarium is equipped with automated cages & devices for behavioral assessments, sample collection and in vivo measurements (ECG, EEG) of the animals. We have a multidisciplinary team of expert scientists who can design a comprehensive project plan to fit any requirement/stage of your research pipeline and for drug discovery and screening.
- Designing and engineering research animal disease models
- Adoptive transfers/transplantation
- In vivo functional screening assays
- End-of-study in vitro assays
Applied StemCell offers gamma-irradiation service for irradiating cell lines (patient-derived, pooled, genome edited/ corrected cell lines) using a Nuclear Regulatory Commission (NRC)-approved Gamma-emission source to irradiate cells at a constant dose rate and to any desired extent of radiation. Irradiated cell line models are novel toolkits for applications such as generation of co-culture models using non-dividing/ feeder cells; to study mechanisms underlying cancer progression; wound healing and inflammatory responses after irradiation; screen drugs for enhancing irradiation sensitivity of tumor cells; and for xenograft tumor models.
Our Cell Line Irradiation Service includes:
- Cell line expansion/ harvesting
- Cell line Irradiation (customized extent of irradiation)
To speed up your research and development; we provide pre-clinical development services (CRO) and support. Our custom services will allow you to focus on the main aspects of your research while we develop the interim paradigms for you.Custom Services Categories
Fibroblasts Immortalization Service
Custom Primary Cell Isolation Service