Stem Cell & Regenerative Neuroscience
Contact
900 Walnut Street, Room 461
Jefferson Hospital for Neuroscience
Philadelphia, PA 19107
Services
The Jefferson Stem Cell and Regenerative Neuroscience Center (JSCRNC) offers iPS cell services including:
- Propagation, expansion, and biobanking of iPS cell lines
- Assistance with differentiation protocol development and validation
- Human iPS cell reprogramming (mRNA, Sendai Virus, Episomal)
- iPS cell characterization
- Consulting with researchers looking to work with iPS cells
- Genome editing (knock in and knock out)
| *Please contact Lorraine Iacovitti, PhD for more information or to begin working with the Center. |
Services
The Center will reprogram patient samples provided by the requesting PI. All material should be IRB approved for iPSC reporgramming and deidentified as set forth in 45 CRF 164.514(b)(2).
| Services | Description | Requirements* | Price |
| Repropgramming fibroblasts using mRNA reprogramming | 1-3 clones per line with full characterization including PCR and IHC for pluripotency, g-band karyotyping, DNA fingerprinting, EB formation for trilineage expression and mycoplasma testing. Feeder-free, Xeno-free, Integration-Free | IRB approved material & Reprogramming Request Sheet |
$5,000-$8,000/line (1-3 clones) |
| Repropgramming blood using mRNA reprogramming | 1-3 clones per line with full characterization including PCR and IHC for pluripotency, g-band karyotyping, DNA fingerprinting, EB formation for trilineage expression, and mycoplasma testing. Feeder-free, Xeno-free, Integration-Free | IRB approved material & Reprogramming Request Sheet |
$5,000-$8,000/line (1-3 clones) |
The Center will thaw and expand IPSC line(s) with approved MTAs for one passage (~2 weeks) and biobank high quality mycoplasma negative cells (x 6 vials).
| Services | Description | Requirements* | Price |
| iPSC Thaw and Expansion | 1 cryovial of iPSC will be thawed into 1 well of a 6-well plate (Geltrex, mTeSR1 media) before being expanded into 6 wells of a 6-well plate. Expect ~2 weeks | cryovial of iPSC, approved MTA & Submission of Application for Use |
$150/line |
| iPSC Cryopreservation | iPSC line(s) cryopreserved in vapor phase of Center's LN2 tanks. 6 vials feeder-free in mTeSR1 + 10% DMSO | Submission of Request Sheet | $150/line |
| + Additional Passages | iPSC line passaging beyond first passage after thaw. | N/A | $50/week |
| + Additional Vials | iPSC line cryopreservation exceeding 6 vials | N/A | $25/vial |
The Center will propogate iPSC line(s) and distribute progeny for requesting laboratories as number of wells/plate(s) as whole colony or monolayer cultures. Requests forongoing passages will incur a $50/week fee for "off weeks" during which no plates of iPSCs are requested.
| Services | Description | Requirements* | Price |
| iPSC Thaw | iPSC line(s) will be thawed into 1 well of a 6-well plate and mainted for a week prior to requests for plates of iPSCs | Submission of Request Sheet | $50/line |
| Plates of iPSCs | iPSC line(s) supplied as whole colony or monolayer (Accutase passaged) Day 5 to Day 7 post-passaging (Geltrex, mTeSR1 media) | Submission of Request Sheet | $50/10cm2 well |
| + Additional Passages | iPSC line(s) maintained as 1 well of a 6-well plate when multiple passages requested | N/A | $50/week |
iPSC differentiation protocol development requires an initial PI consultation with Dr. Iacovitti to discuss project goals and desired lineage(s). PI is responsible for researching current methods for differentiating desired lineage(s) and should bring reference material with them to initial consultation. Once a differentiation protocol is agreed upon, the Stem Cell Center will begin validating the protocol. Expect ~3 months for protocol validation. PI will receive a detailed written protocol to replicate iPSC differentiation in their laboratory. No iPSC progeny, differentiated material, or reagents will be transferred.
| Services | Description | Requirements* | Price |
| iPSC Differentiation Protocol Development | Validation of desired differentiated iPSC phenotype up to the point of "state of the art" in the field | Cell Culture Experience Submission of Application for Use |
$5,000 |
| + Additional Assistance & Troubleshooting | Session with Dr. Iacovitti and Center personnel to discuss iPSC differentiation protocol and troubleshoot issues | N/A | $200/session |
Disclaimer: Current iPSC differentiation methods are not 100% efficient, so a pure culture can not be guaranteed. The Stem Cell Center will work to achieve a culture that is comparable to other published work, but will not be responsible for iPSC differentiation beyond “state of the art” at the time of the request.
Our center will generate genome edited iPSC lines using the CRISPR-Cas9 technique for cell-based therapy or regenerative research. Expect 3-6months to get the verified clones. Point mutation G>A was fixed in the isogenic line (A, top) and tdTomato expression was observed when one iPSC reporter line was differentiated toward a dopaminergic neuron lineage (B).
| Services | Description | Requirements* | Price |
| CRISPR-Cas9 mediated knock out | KO of your gene of interest in iPSC lines | Submission of Request Sheet | To be determined according to the project |
| CRISPR-Cas9 mediated mini knock-in | Point mutations or other small mutations can be corrected or generated in iPSC lines | Submission of Request Sheet | To be determined according to the project |
| CRISPR-Cas9 mediated maxi knock-in | A reporter gene can be inserted into your gene of interest in iPSC lines | Submission of Request Sheet | To be determined according to the project |