iPSC have been generated by reprogramming Lymphoblastoid Cell Lines (Blood. 2011 Aug 18;118(7):1797-800, Cell Cycle. 2011 Sep 1;10(17):2840-4, Blood. 2011 Aug 18;118(7):1801-5) Therefore we will pilot 4 methods to generate iPSC from lymphoblastoid cell lines generated in the ALSPAC laboratory.

There are four objectives for the pilot study:

1. Establish reprogramming of lymphoblastoid cell lines using 4 different methods

2. Identify and expand a number of prospective iPSC lines from the six EBV lines supplied by ALSPAC

3. Characterise the resulting iPSC using normal criteria used to asses pluripotency

4. Analyse the possible removal of the EBV genome from iPSC lines during derivation / extended culture

SAMPLE REQUEST

Six lymphoblastoid cell line samples will be chosen and distributed to the 4 laboratories. These will be selected using following criteria

3 male, 3 female

all with UK10K data

all ARIES cases

LCL's established from PBL obtained at 9yr clinic

PBL also available from F17 clinic

Methods to be used are detailed in appendix 1b, in summary groups will use the following

Lyle Armstrong, Majlinda Lako, University of Newcastle

Sendai virus reprogramming of lymphoblastoid cell lines

Maeve Caldwell, James Uney, University of Bristol

Reprogram the LCLs into iPS cells using the lentiviral system

Kevin Eggan, Harvard University

Generate iPSC from provided LCL lines using episomal plasmid based methods

Amanda Fisher, MRC Clinical Science Centre, London

heterokaryon iPSC generation system