iPS Cell Service Request Form
Email: iPSCORE@ucdenver.edu Web Site: medschool.ucdenver.edu/iPS
Investigator:
Contact Email:
Speedtype:
Date:
Phone:
Speedtype:
Specimen: Human ○ Mouse ○ Other:
List known diseases of human donor cells:
iPS Cell Services (Step 1)
Price
Isolation of Primary Fibroblast from Skin Biopsy or other Sources for iPS
Generation: expanding and freezing of up to 5 vials of cells
Expansion of Primary Cells for Transduction: expanding and freezing of up to 5 vials of
$607
cells
Quantity Subtotal
$451
Transduction of Primary Cells (Step 2)
Reprogramming System1: Non-Integrating (Sendi Virus) Transduction of Primary Cells: up to
5 iPS cell clones; includes mycoplasma testing and RT-PCR for 3 pluripotency markers (Oct4,
Nanog, SOX2)
Reprogramming System2: Integrating Lenti-Virus Transduction of Primary Cells: up to 5 iPS
cell clones; includes mycoplasma testing and RT-PCR for 3 pluripotency markers (Oct4, Nanog,
SOX2)
$6,000
$6,000
Validation and Characterization Services (Step 3)
Immunocytochemistry Staining for Pluripotency Markers Present in
Undifferentiated iPS cells: Cells are tested for the presence of Oct4, Nanog, TRA-160, SSEA3,
$1,084
or SSEA4
QRT-PCR for Pluripotency Markers: (Oct4, Nanog, Sox2) for quantification and
comparison of marker expression in iPSC and hES Cells. Other markers upon request.
RT-PCR for Pluripotency Markers: (Oct4, Nanog, SOX2) non-quantitative evaluation of
stem cell marker expression. Other markers upon request.
Karyotype Analysis/clone: Assess for normal chromosomal numbers, sex determination and
chromosomal aberrations
Embryoid Body (EB) Differentiation Assay: Differentiation potential to form the 3 germ
layers by RT-PCR or ICC staining for the following differentiation markers: mesoderm (SMA),
endoderm (anti-Tuj1) and ectoderm (AFP)
Mycoplasma Testing of iPSC lines
$823
$701
$1,139
$1,142
$316
Additional Services (Step 4)
Embroid Body (EB) Formation only
iPS Cell line Lenti/Sendai (Core Generated; price per vial)
Clone Expansion (10 Vials per iPS Cell line)
Other Pluripotency Markers by QRTPCR and RTPCR
Distribution Services of iPS Cell lines
$94
$650
$659
Inquire
Inquire
Subtotal
Administration Fees (Step 5)
Internal Customers – within the University of Colorado
External US Customers – within the United States
External International Customers – outside of the United States
For-Profit Customers – within or outside of the United States
NA
5%
10%
15%
Total
Description and Explanation of Services
Step 1: Isolation of Fibroblasts: The core will establish fibroblast cells from skin biopsy or expand fibroblast cells provided to us.
Fibroblast cells must be at low passage and in active proliferation without signs of senescence for good reprogramming.
Additionally, fibroblast will be tested for mycoplasma infections and frozen stocks will be established. This step should take
approximately 1 month.
Step 2: Transduction of Primary Cells: The core will transduce fibroblasts to generate iPS cells using either a Sendai virus based
vector system (non-integrating vector system) or a lenti-virus based vector system (integrating vector system with excisable
reprogramming cassette). We will isolate 5 iPS cell colonies per experiment; this should provide the investigator with several
karyotypically normal iPS cell lines. At this point, 5 frozen vials are available to the investigator. This step should take
approximately 2 months.
*Note that the core can also utilize other primary cell types other than fibroblast and from different species for the generation
of iPSC. Please inquire about requirements.
Step 3: Validations and Characterization Services: The Core can provide a variety of characterization services. We recommend
at a minimum the following characterization services: full karyotyping analysis, QRT-PCR for pluripotency expression, and
embryoid body (EB) formation to assess differentiation potential of the iPS cells. Other characterization services listed in the
table above are strongly recommended. Other pluripotency markers are available for screening by QRTPCR and RTPCR under
additional services upon request. Data will be provided to the investigator upon completion of all services. This step should take
approximately 1 month.
Step 4: Additional Services: Once the established iPS cell line has been validated and characterized, the core will expand and
generate 10 vials of frozen stock per line for the investigator. Upon request, the iPS Core can serve as a repository and provide
distribution services for all in-house generated iPS cell lines (inquire about conditions for this service).
Step 5: Administration Fees: These fees cover all communications, paperwork, data entry, and handling of shipments.