• iPSC Generation from Patient Samples

iPSC Generation Services

Let the iPSC expert generate high-quality, induced pluripotent stem cell (iPSC) lines and derived physiologically relevant cell line models for your basic research, drug discovery, drug screening, and preclinical cell regeneration projects:

  • Highly optimized protocols with high reprogramming efficiency (>95% success rate)
  • From healthy/diseased samples
  • iPSC generation from PBMCs, fibroblast, HSC, MSCs, CD34+ cord blood, urine, and more
  • integration-free (episomal/ mRNA/ viral-based) or retroviral reprogramming
  • Feeder-free protocols; optional feeder-dependent protocols available
  • iPSCs characterized for morphology and pluripotency markers. Additional characterization such as G-banding, RT-PCR, STR profiling, directed-differentiation are also available.   
iPSC Generation Services Categories

iPSC Generation
from Patient Samples

Efficient iPSC generation from healthy/ disease patient samples for cell line/ disease modeling, drug discovery and drug/ neurotoxicity screening.

iPSC Generation
from Patient Samples

Allogenic and Immunocompatible
iPSC Generation

Full service for HLA-matched allogenic iPSC generation and CRISPR gene editing to engineer HLA-knockouts for developing non-immunogenic iPSCs.

Allogenic and Immunocompatible
iPSC Generation

Support Materials
FAQs
What will customers need to provide for iPSC generation from patient fibroblasts?
What is the minimum numbers of cells needed for iPSC generation from PBMCs?
Technical Details
 
 
schematic-ipsc-generation-services
 

Induced Pluripotent stem cells (iPSCs) are a type of pluripotent stem cells that can be reprogrammed from adult somatic cells derived from healthy or diseased patient samples such as skin biopsy (fibroblasts), peripheral blood mononuclear cells (PBMCs), hematopoietic stem cells (HSCs), mesenchymal stem cells (MSCs), urine and more. Similar to embryonic stem cells (ESCs), the iPSCs have the ability to propagate indefinitely and to differentiate into any type of cell in the body. They represent the genetic and epigenetic characteristics of a patient better and circumvent the ethical and sourcing issues associated with embryonic stem cells (ESCs). The iPSCs can also be genome engineered which has been a stumbling block for developing research models using primary cells lines. Genome-edited iPSCs can then be differentiated into various somatic lineages and can provide a limitless supply of patient-derived cell lines from the same source tissue. The iPSCs are a very valuable resource for developing clinically relevant in vitro disease models for basic research, drug discovery and drug screening; precision medicine, and cell-based therapies in regenerative and transplantation medicine. Genome engineered iPSCs have been successfully to develop universal iPSCs/ non-immunogenic iPSCs for allogenic transplantation, CAR-T and other immunotherapies.

Applied StemCell has played a crucial role in the iPSC area. We have generated several hundred iPSC lines for scientists in academia and industry from a variety of patient samples (healthy/disease) as well as large scale generation for big biopharmaceutical companies. We can work with iPSC banks and other organizations that require large-scale generation of iPSC lines using highly optimized, standardized, and efficient methods. We can reprogram iPSCs with a very high success rate (>95%) using non-integrating reprogramming factors (such as episomal, non-retroviral virus, mRNA) from a variety of starting patient samples and using feeder-free (and xeno-free) protocols, and with a fast turnaround time (2-3 months):

Skin (dermal) fibroblast

Hematopoietic stem cells (HSCs)

Urine

Peripheral blood mononuclear cells (PBMCs)

Mesenchymal stem cells (MSCs)

Hair follicles

Cord blood cells (CD34+)

Adipose-derived stem cells (ADSCs)

Human umbilical vein endothelial cells (HUVEC)

If you require retroviral method for iPSC generation and/or feeder-dependent protocols, please discuss your specific requirements with our team of experts.

Got your iPSCs? Make it a one-source-shopping experience! We provide a complete range of iPSC-related services for your projects with our services for downstream expanded characterization, or genetically engineer iPSCs using CRISPR/Cas9; and differentiation to somatic lineages such as neuronal lineage cells, microglia, T cells, NK cells, retinal pigment epithelium (RPE), cardiomyocyte, hepatocytes, and more.

Applied StemCell is a member of the National Institute of Standards and Technology (NIST) Genome Editing Consortium and brings to the consortium its expertise in genome editing (CRISPR/Cas9 and proprietary TARGATT™) and induced pluripotent stem cell technologies for the advancement of drug discovery and therapeutic applications.

Workflow and Timeline:

Service Time
1. Pathogen test
1-2 weeks      
2. Cell recovery, expansion, and banking      3-4 weeks    
3. Transfection with vectors and iPSC generation   
3-4 weeks    
4. Colony picking, expansion, and cryopreservation    
3-4 weeks    
5. iPSC characterization (standard)* 
2-3 weeks    
Total
2-3 months

* more detailed characterization options are available upon request

Service details:

  • Please provide three (3) vials with 1 x 10^6 cells/vial from patient samples 

Deliverables:

  • Two (2) iPSC clones with two (2) vials per clone at 1 x 10^6 cells/ vial
  • Clones are characterized for three (3) pluripotency markers: OCT4, SOX2, SSEA4, TRA-1-60, TRA-1-81

Applied StemCell can also genetically engineer iPSCs using CRISPR/Cas9 and can provide expanded characterization and differentiation to somatic lineages such as neuronal lineage cells, microglia, T cells, NK cells, retinal pigment epithelium (RPE), cardiomyocyte, hepatocytes, and more.

 

Case Studies

Case Study: Generation of Human iPSCs from Patient-derived Skin Fibroblasts

Goal: To generate an iPSC line from patient derived dermal fibroblasts.

The dermal fibroblasts were obtained by skin biopsy from a patient with a rare disorder. The fibroblasts were plated, transfected via electroporation and cultured with ASC’s proprietary iPSC generation media until iPSC colonies appeared. Five high quality candidate colonies were expanded and characterized by immunocytochemical staining using primary antibodies for: OCT4, SOX2, SSEA4, TRA-1-60 and TRA-1-81) and alkaline phosphatase staining. All five clones stained positive for various pluripotency markers tests as well as for alkaline phosphatase (Figure1).  

CASESTUDY-SERVICE-StemCell-iPSC-Generation-disease-fibroblast

Figure 1. Human iPSCs were generated from patient-derived dermal fibroblasts, using feeder-free, proprietary protocols. The reprogrammed iPSC clones were characterized by pluripotency marker staining for OCT4, SOX2, SSEA4, TRA-1-60, TRA-1-81, and alkaline phosphatase (AP).

Publications
  • Jang, Y., Choi, J., Park, N., Kang, J., Kim, M., Kim, Y., & Ju, J. H. (2019). Development of immunocompatible pluripotent stem cells via CRISPR-based human leukocyte antigen engineering. Experimental & Molecular Medicine, 51(1), 3.
  • Ilic, D. (2019). Latest developments in the field of stem cell research and regenerative medicine compiled from publicly available information and press releases from nonacademic institutions in October 2018. Regenerative medicine, 14(2), 85-92.
  • Allende, M. L., Cook, E. K., Larman, B. C., Nugent, A., Brady, J. M., Golebiowski, D., ... & Proia, R. L. (2018). Cerebral organoids derived from Sandhoff disease induced pluripotent stem cells exhibit impaired neurodifferentiation. Journal of Lipid Research, jlr-M081323.
  • Field, A. R., Jacobs, F. M., Fiddes, I. T., Phillips, A. P., Reyes-Ortiz, A. M., LaMontagne, E., ... & Hauessler, M. (2019). Structurally Conserved Primate LncRNAs Are Transiently Expressed during Human Cortical Differentiation and Influence Cell-Type-Specific Genes. Stem cell reports.
Have Questions?

An Applied StemCell technical expert is happy to help, contact us today!