• Master cGMP-Grade iPS Cell Line
    • iPSC Expansion and Banking Master & Working Cell Banks
    • iPSC Reprogramming, Genome Editing & Differentiation
    Master cGMP-Grade iPS Cell Line

GMP Grade iPSC Services & Products

Leading iPSC CDMO, Ready-to-use iPSC GMP Master Cell Bank & GMP Cell Therapy Services

ASC received its Drug Manufacturing License from the California Food and Drug Branch in 2021. Since then, iPSC processes have been established for cell banking and iPSC reprogramming, genome editing (CRISPR and TARGATT™), and differentiation that are compliant with FDA (US) and EMA (EU) regulations. As a CRO/CDMO committed to providing high-quality services and products, we follow quality control protocols based on ICH-Q5A/D guidelines for the development of safe iPSC and iPSC-derived products in our ISO Class 5 and 7 (Class 10,000 and 100) clean room.

In addition to our cGMP-compliant iPSC generation service, we offer well-characterized, ready-to-use GMP-grade iPSCs (Cord Blood CD34+, Male) and cGMP-Compliant TARGATTTM Master iPSCs that can be engineered to express almost any gene of interest (e.g., CAR genes). The genome-edited cells can be further differentiated into various cell types, including NK cells, T cells neurons, RPE cells, and more. Our team of experts can even work with customer-provided cell lines. We currently offer:

Current Good Manufacturing Practice (cGMP):

Regulations ASC complies with as a cell processing GMP facility <Located in CA, USA> 

  • 21 CFR Part 210
    • Current Good Manufacturing Practice in Manufacturing, Processing, Packing or Holding of Drugs; General
  • 21 CFR Part 211
    • Current Good Manufacturing Practice for Finished Pharmaceuticals
  • 21 CFR Part 1271
    • Human Cells, Tissues, and Cellular and Tissue-Based Products
    • Current Good Tissue Practice

1. GMP Grade iPSCs

  1. cGMP-Grade iPSC Line: The cells are well characterized and can be genetically modified and further differentiated to various cell types.
    • Source cell type: CD34+ Cord Blood, male
    • Reprogramming method: Episomal
    • Master cell bank or working cell bank available
    • Drug master file with US FDA
    • Matching Research iPSC Line is Available

Product QC:

  • Sterility/mycoplasma
  • Identity: SNP Trace
  • Pluripotency: image, AP stain, Oct4/TRA-1-60 stain, differentiation in vitro to 3 germ layers
  • Genetic stability: karyotyping by G-banding
  • Viability
  1. cGMP-Compliant TARGATTTM Master iPSCs: Ready to insert your GOI. High-quality iPSCs with a pre-engineered landing pad for site-specific, unidirectional gene insertion at a safe harbor locus mediated by TARGATTTM
    • Engineered from ASC’s GMP grade iPSCs using TARGATTTM (No CRISPR)
      • GMP SOP
    • All reagents are CGT grade (animal, serum-free)
    • Documentation and QC/QA follow GMP SOP
    • Certificate of Analysis provided upon request
    • All testing follows GMP requirements
    • License-ready  

2. GMP iPSC Services

GMP iPSCs

 

iPSC GMP SERVICES: iPSC Generation, Expansion, and Banking

cGMP iPSC Expansion and Banking

 

Number of Vials

Cell Count

Other Deliverables

Master Cell Bank

50-200 vials

1x10^6 cells/vial

QC and CoA

Working Cell Bank

50-200 vials

1x10^6 cells/vial

QC and CoA

*Stocking of Frozen Cells (Annual Storage): 24/7 monitoring

 

Customized iPSC Generation (Episomal Method)

 

Number of Clones

Number of Vials

Cell Count

Other Deliverables

Seed Bank

3 Clones

10 vials

1x10^6 cells/vial

QC Characterization

and CoA

GMP iPSCs

GMP SERVICES: iPSC Gene Editing

With 13+ years of stem cell & genome editing experience, ASC offers high-quality gene editing services using the latest technology, CRISPR and TARGATTTM. For all CRISPR, fee-for service gene editing projects, the customer must obtain a CRISPR license. With ASC’s TARGATTTM technology, customers can avoid having to obtain additional licensing and only need a single license from ASC!

iPSC Gene Editing

Service

Details

CRISPR/Cas9 Gene Editing

  • Customer needs to obtain CRISPR license
  • ASC performs fee-for-service gene editing

TARGATT™ Gene Editing

  • License terms already in place

 GMP-like Plasmid Production For Gene Editing

  • QC: purity, sequence, residual host DNA/protein/RNA test, etc.

In-Process QC:

  • Reagent QC (plasmid map/sequence)
  • Validation (cell, gRNA, integrase)
  • Transfection pool genotyping
  • Clone genotyping

TARGATTTM Site-Specific Knockin Technology

Unlike other gene editing companies, ASC has a unique gene-editing technology, TARGATTTM, that enables site-specific, irreversible gene insertion at a safe-harbor locus, H11 or ASC2. The unidirectional gene insertion at an active, intergenic locus does not disrupt other internal genes. TARGATTTM works well in non-dividing cells and permits single-copy insertion of large fragments up to 20kb. ASC has observed high integration efficiencies and medium to high protein expression in stable cells.

The NIH awarded Applied StemCell with a $2M SBIR grant for the improvement of its hTARGATTTM technology for human gene and cell therapy. According to the NIH ASC’s proprietary gene editing technology

 

 “could efficiently and safely treat more than ten thousand monogenic human diseases.”

                                                        - NIH grant review group summary statement

GMP iPSCs

GMP SERVICES: iPSC DIFFERENTIATION

Integration-free protocols to differentiate iPSCs to various committed somatic lineages, including NK cells, T cells, RPE cells, neurons, and many others.

 

iPSC Differentiation

Service

Details

iPSC Differentiation: Customer provides protocol

  • Tech transfer
  • Engineering run/scale up
  • cGMP production

iPSC Differentiation: ASC’s SOP

  • Engineering run/scale up
  • cGMP production

GMP iPSCs

GMP SERVICES: iPSC DIFFERENTIATION TO COMMITED SOMATIC LINEAGES

GMP iPSC Differentiation

GMP iPSC Differentiation, differentiated cell types

 

GMP SERVICES: iPSC DIFFERENTIATION TO NK CELLS (iNK)

Characterization of the ASE-9708 NK Cells

Natural Killer Cells - ASE-9708

Figure 1. Flow cytometry analysis of ASE-9708 iPSC-derived NK cells for NK cell biomarkers. Cryopreserved NKs, differentiated from Applied StemCell’s control iPSC line, ASE-9211 were recovered in NK culture media. The cells were stained with NK cell markers, CD45 and CD56 at day 2. Left: Isotype control antibodies. Right: CD45/CD56.

GMP SERVICES: iPSC DIFFERENTIATION TO RPE CELLS

Characterization of the ASE-9710 RPE Cells

Figure 2. Immunostaining of ASE-9710 iPSC-derived Retinal Pigment Epithelium Cells for retinal pigment
epithelium biomarkers.
Cryopreserved RPE cells, differentiated from Applied StemCell’s control iPSC line, ASE9211 were recovered in RPE culture media. The cells were stained with RPE markers, PMEL1, MITF, ZO-1 and
RPE65.

TARGATTTM Master iPSC Platform for Allogeneic Products

The comprehensive TARGATTTM Master iPSC platform permits the safe and efficient insertion of any gene of interest at a safe harbor locus in the TARGATTTM Master iPSCs. This process is facilitated by TARGATTTM integrase, and the engineered cells can be further differentiated to various cells types.  

 

  1. TARGATTTM Master iPSCs

Pluripotent stem cells with a pre-engineered landing pad in a safe genomic locus for repeated, fast, and efficient gene knock-in by TARGATTTM integrase

  • Engineered from ASC’s GMP grade iPSC using TARGATT™ technology only (no CRISPR)
  • All reagents are CGT grade (animal, serum-free)
  • Follow GMP SOPs
  • Documentation and QC/QA follow GMP SOP
  • Certificate of Analysis provided
  • All testing follows GMP requirement
  1. iPSC Differentiation

For the creation of unlimited sources of off-the-shelf allogenic therapeutic cells.

  • CAR-T or CAR-NK cells (no need for lentiviral transfection), neurons, or other cell types

GMP iPSCs

TARGATT™ iPSC-iNK Platform in CAR-iNK Therapy

Current Unmet Needs Using CAR-T/NK Therapy

  1. Autologous CAT-NK/T products can only be used by a single patient and it is expensive
  2. Random CAR gene insertion caused by lentivirus or transposon methods is unsafe
  3. CRISPR limits the payload size and has a low efficiency
  4. Current lentivirus manufacturing of CAR-NK/T products

TARGATT™ iPSC-iNK in CAR-NK Therapy

Our TARGATTTM system can be used to knock-in your CAR gene. ASC’s experts can confirm your iPS cells express CAR and differentiate your CAR-positive cells into NK cells or T cells.

 

CAR Knock-In

  • CAR was inserted at >40% efficiency
  • 10x better than CRISPR/Cas
  • CAR-iPSC express CAR
  • Transfection done by lipofectamine,
    eliminating viral manufacturing

 

CAR positive iPSC line express CAR

Figure 3: Flow cytometry analysis of iPSC lines screened for surface expression of CAR. Monoclonal antibodies labeled with a fluorescent molecule were used for screening. During the screening process, an iPSC line that expressed CAR was identified. (Images: Left: CAR negative iPSCs; Right: CAR positive iPSCs)

CAR positive iPSC-iNK cells express CAR

Figure 4: qRT-PCR data for expression levels at each stage of the iPSC to NK cell differentiation process.

Figure 5: qRT-PCR data for OCT4 and CD56 levels throughout the differentiation process, including the final CD56 stage.

GMP iPSCs

Products and Services
Technical Details

GMP Facility

ASC's GMP facility is set for iPSC generation, expansion, gene editing, and differentiation as well as general cell banking processes. ASC’s GMP facility includes manufacturing space, the storage for raw and finished products, and support lab areas. ASC’s GMP facility operates under the guidelines established by the CFR (Code of Federal Regulations) Title 21, Parts 1271 (Current Good Manufacturing for Human Cells, Tissues, and Cellular and Tissue-Based Products).

Application Notes

Applications in Cell Therapy

The off-the-shelf master iPSCs have the potential to revolutionize immunotherapy. The engineered human induced pluripotent stem cells with a landing pad at a genomic safe-harbor locus can be used to insert a therapeutic gene of interest to generate an iPSCs that carry a single copy of the gene needed for therapeutic studies.

Universal Modifiable iPSC in CAR-NK Therapy

Since ASC’s Master iPSC line holds a TARGATTTM site at a safe-harbor locus, a single copy of a gene of interest such as CAR can be inserted into the genome. ASC could generate an iPSC line with a specific CAR, and use its integration-free method to differentiate the iPSCs with CAR to a committed cell type such as natural killer (NK) cells. With the rise in CAR-NK and CAR-T therapies, ASC’s TARGATTTM and iPSC differentiation capabilities offer an alternative, effective method for the development of novel CAR-NK cell therapies.

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