Patient sample processing: isolation, expansion, and banking
Skin biopsies
Biopsies should be collected into a tube with DMEM + Pen/Strep, which can be provided to on-campus users. The biopsies should be refrigerated after collection and transported to the GESC on ice. Do NOT freeze or place the samples on dry ice. If shipped to the GESC, the samples should be packed with blue ice and shipped overnight.
Urine
The urine samples have to be processed within an hour of collection. It is helpful to dilute the urine sample(s) immediately with 1 volume of ice-cold DMEM (or other base media) after collection. The sample(s) should be brought to us on ice as soon as possible. It is always helpful to let us know ahead of time that samples will be brought over so that we can coat the plates beforehand.
Whole blood
5-10 mL of whole blood should be collected into purple (EDTA) or green (Heparin) top collection tubes to prevent clotting. The samples should be kept in the fridge until transportation. Blood samples processed within 24 hours give us the best yield of erythroid progenitor cells and the best outcome from reprogramming. It is possible to recover erythroid progenitor cells from clotted blood samples with a lower yield.
Hands-on, on-demand training on iPSC culturing
Stem cells are cultured very differently from cancer cells. For researchers new to stem cell work, a brief training saves money, time and frustration. The training spans a week with about 5 hours of total hands-on time. It starts on a Monday and by Friday of the same week, you will have learned to prepare plates and medium, maintaining and splitting iPSCs, recognizing and cleaning up differentiated cells in culture, freezing down iPSCs. You will also bring back with you a live culture and/or cryo vials if you wish. Your trainer will continue to be your source of support on your iPSC techniques after the training. To schedule training, please contact Amber Neilson, aneilson@wustl.edu
Reprogramming of primary cells into induced
pluripotent stem cells (iPSCs)
Starting cells
Patient samples processed at the GESC or sent by investigators
Method
We use the CytoTune 2.0 kit for reprogramming, which is a Sendai virus cocktail. Sendai virus is a non-integrating RNA virus
General timeline
6-8 weeks. We deliver up to three clones per patient sample.
iPSC QC assays
Karyotyping through Cytogenetics
Turnaround time is on average 2-4 weeks
Pluripotent staining
Turnaround time is on average 1-2 weeks
Embryoid body formation
Turnaround time is on average 3-4 weeks
iPSC Differentiation
Currently, the offer includes the following cell types:
- Cardiomyocytes
- Neural progenitors
- Neural stem cells
- Hepatocytes
- Hematopoietic stem cells
- Macrophages
- Astrocytes
- Motor neurons
- Sensory neurons
- Adipocytes
- Monocytes
If the cell type of interest is not listed, please inquire
To submit Patient sample processing, Reprogramming and Differentiation requests, go here, if you are on WashU network. Instructions can be found here. If you are a WU employee working off site, you have to connect to the WashU network via VPN. For non-WU users, please contact us.
- 2023 JIT Voucher Program | PCEN | Washington University in St. Louis
- Just-In-Time (JIT) Core Usage Funding Program | Clinical & Translational Sciences | Washington University in St. Louis
- Shared Resources (Cores) – Research – Siteman Cancer Center
- CDI Pediatric Disease Mouse Models Core (PDMMC), up to 70% subsidy towards eligible mouse models. Please contact Dr. Jeffrey Miner for details: jeffminer@wustl.edu.
- The Rheumatic Diseases Research Resource Based Center (RDRRC) offers reimbursement for GEiC services, up to $4000 for junior faculty and $1000 for other faculty with a brief, easy, and quick application process. Visit them here.