Reprogramming


The core uses a patent-pending, modified mRNA/miRNA-based method to reprogram dermal fibroblasts, urine derived renal epithelial cells, and amniotic fluid derived stem cells into induced pluripotent stem cells (iPSCs). This cutting-edge technology was developed by the co-directors of the core, Drs. Kogut and Bilousova (High-efficiency RNA-based reprogramming of human primary fibroblasts. Nature Communications. (2018). https://doi.org/10.1038/s41467-018-03190-3). It provides the following significant advantages over many other cell reprogramming approaches:

  • non-integrative
  • ultra-high yield (up to 90%)
  • cost effective

reprogramming 

We also offer the integration-free episome-based reprogramming for the cells that are resistant to RNA-based reprogramming, such as peripheral blood mononuclear cells (PBMCs).

 

We provide up to 4 iPSC clones per reprogrammed cell line (2 vials per clone).


Fibroblast Reprogramming

 

Primary fibroblast lines provided by a customer, or isolated by the core from provided skin biopsies, are reprogrammed into induced pluripotent stem cells (iPSCs) using the RNA-based approach. If desired, a complete characterization profile can be provided post reprogramming. 

ServiceTime
Fibroblast Isolation From Skin Biopsy2 weeks
Fibroblast Reprogramming2-3 weeks
iPSC culturing and expansion2 weeks
Mycoplasma test1 week
Pluripotency Staining1 week
Karyotyping4 weeks

Renal Cell Reprogramming

The core personnel is skilled in developing primary renal epithelial cell (REC) lines from human urine specimens and reprogramming REC lines into iPSCs. Please contact us for more information.

 

PBMC Reprogramming

 

The core reprograms PBMCs using the following episomal plasmids:

 

  • pEV-SFFV-Oct4-2A-Sox2 Vector
  • pEV-SFFV-BCL-XL Vector
  • pEV-SFFV-Myc Vector
  • pEV-SFFV-KLF4 Vector

 

The resulting iPSC clones are expanded for up to 6-10 passages to eliminate the residual episomal vectors before the cells are released to a customer. PBMC reprogramming and iPSC expansion to eliminate episomes take approximately 3 months.

Note: a copy of approved IRB protocol(s) must be provided by customers before reprogramming patient-derived cells

Gates Institute

CU Anschutz

Anschutz Health Sciences Building

1890 N Revere Ct

Mailstop L606

Aurora, CO 80045

303-724-1494


Gates Biomanufacturing Facility

12635 East Montview Blvd

Suite 380

Aurora, CO 80045

720-576-2722


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