Primary cell isolation, banking, mycoplasma testing

​Note: It is the customer’s responsibility that the donor material is taken with proper informed consent. A template informed consent form is available upon request.

The Leiden hiPSC Centre offers isolation of primary cells from the following tissue sources:

Skin (punch biopsy, minimal 4 mm):

Fresh punch biopsies are preferred; storage in PBS at 4°C for up to 14 days is possible. Skin fibroblasts from the dermis are expanded, tested for the absence of mycoplasma and cryopreserved at 2 early passages.

Note: In the Netherlands, taking punch biopsies from minors is not permitted.

Peripheral blood (15 ml in sodium heparin tubes)

Fresh blood is preferred; storage for a maximum of 24 hours at RT is possible. Peripheral blood mononuclear cells (PBMCs) are isolated by Ficoll gradient, tested for the absence of mycoplasma and cryopreserved. For reprogramming erythroblasts after selective expansion from PBMCs will be used.

Peripheral blood (80 ml in sodium heparin tubes)

Only fresh blood. Blood outgrowth endothelial cells (BOECs) are isolated, tested for the absence of mycoplasma and cryopreserved at 2 early passages.

Note: Due to the scarcity of BOECs in blood, the success rate for their isolation is low. In the Netherlands, the maximum blood volume taken from minors is 40 ml.

Urine (150 ml minimum)

Only fresh urine. Renal epithelial cells are isolated, expanded, tested for the absence of mycoplasma and cryopreserved at 2 early passages.

Milk teeth, fresh

Dental pulp cells are isolated, expanded, tested for the absence of mycoplasma and cryopreserved at 2 early passages.

Note: Compared to other donor material dental pulp cells have an increased risk for failure of isolation due to frequent cell culture contaminations.

Additional cell types, e.g. from surgical waste material

Upon request.

After completion of reprogramming, any leftover cryovials with primary cells are transferred to the customer.