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

Submit Patient Sample Processing, Reprogramming and Differentiation Requests

 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 or a non-WU user, you have to connect to the WashU network via VPN. For non-WU users, if this is the first time to use the site, you have to request a guest WUSTL key for VPN first by clicking here. Please use your institutional email address for the request. And if you are not the PI, please specify both your institution and lab under “Organization”. Sponsor’s email address: x.cui@wustl.edu. You will receive your credentials via email once the request is approved. And the credentials will only allow you to access this specific server. Please use Contact us to report any issues.  

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