NKI GEMM-ESC archive
The Netherlands Cancer Institute (NKI) has created a unique archive of embryonic stem cells (ESCs) derived from validated genetically engineered mouse models (GEMM) of cancer. These GEMM-ESCs, with often multiple modified alleles, form the basis for further genetic engineering either by Flp-recombinase mediated integration, gene targeting or Crispr/Cas9 to allow for the evaluation of altered target gene expression in a spontaneous tumor model.
The culture and manipulation of GEMM-ESC clones is performed entirely under feeder- and serum-free conditions using the defined N2B27 medium with LIF and the two inhibitors (2i), CHIR99021 and PD0325901, as originally described by the group of Austin Smith, Cambridge, UK. Once the desired genetic alteration is introduced in a GEMM-ESC clone, chimeric mice can be produced using the classic ESC injection protocols that are routinely applied in transgenic facilities. Interestingly, the chimeric mice can be directly used to monitor tumor development as they contain the same genetic alterations as the original GEMM including the altered target gene expression. This GEMM-ESC approach allows for rapid and scalable target gene validation in vivo.
Currently the GEMM-ESC archive contains seven models; four models breast cancer, one for small cell lung cancer (SCLC), one for non-small cell lung cancer (NSCLC) and a model of malignant mesothelioma. A detailed protocol can be found at:
Nature Protocols 2015 Oct 15. Using the GEMM-ESC strategy to study gene function in mouse models. Huijbers IJ, Del Bravo J, Bin Ali R, Pritchard C, Braumuller TM, van Miltenburg MH, Henneman L, Michalak EM, Berns A & Jonkers J.
The NKI GEMM-ESC archive is housed at the Mouse Clinic for Cancer and Aging (MCCA) in Amsterdam, the Netherlands. GEMM-ESCs clones are sent under a Material Transfer Agreement (MTA). Shipping costs are for the receiving party and a 500 EUR handling fee is charged per clone
Direct contact for scientific enquiries and clone distribution is:
Paul Krimpenfort PhD
The Netherlands Cancer Institute
Plesmanlaan 121
1066 CX Amsterdam
The Netherlands
+31 (0)20 512 1992
Mouse Clinic for Cancer and Aging (MCCA)