Module 5: Development of Pluripotency Reporter Lines for Facile Screening of Bioreactor Conditions

Putative hESC pluripotency  reporter line (Photograph, Mirabelle Ho)

Module Leader Dr Andrew Laslett
Host Organisation CSIRO

Module description

The objective of this module will be to create human embryonic stem cell (hESC) pluripotency reporter lines for use in the development of bioreactors for the safe expansion of hESC.

These reporter lines, once constructed, will be invaluable tools for the high-throughput screening of bioreactor conditions to ascertain the optimal conditions for the expansion of hESC and also as a tool for the detection of contaminating pluripotent cells in bioreactors being used to expand differentiated cell types made from hESC. In addition these reporter lines will be valuable tools for the rapid assessment of media components to help in the construction of novel defined media for the maintenance of hESC.

Aims

The overall hypothesis to be tested for this portion of the collaborative stream is that bioreactors can be used to safely expand hESC.  This module aims to provide essential tools to allow for the rapid development of bioreactor technologies and aims to:

  1. generate and characterise hESC pluripotency reporter lines using a lentiviral expression system
  2. generate and characterise stably transfected hESC pluripotency reporter lines utilising targeted knock-in constructs.  

Module Leader biography

Andrew and his team joined CSIRO Molecular Health Technologies in August 2009 and they remain based at the Australian Stem Cell Centre (ASCC) in Clayton. Prior to this he was a Senior Scientist and Group Leader of the Human Embryonic Stem Cell Technology Laboratory at the ASCC.  Dr Laslett’s research compares human embryonic stem cells to human induced pluripotent stem cells and is focused on exploiting the basic biology of these cell types to create novel tools that enhance pluripotent cell research translation. He leads an independent program as well as having significant national and international collaborations.

In September 2007, Dr Laslett was elected as a Board Member and Director of the Australian Society of Medical Research (ASMR). Dr Laslett’s research is supported by the Australian Stem Cell Centre, the NHMRC, the Victoria California Stem Cell Alliance (Victorian Government and California Institute for Regenerative Medicine) and the NSW / Victorian Government Stem Cell Research Grant Program.

Contact details

 E-mail  andrew.laslett@csiro.au 
 Phone  +61 3 9271 1133

Selected publications

  1. LASLETT AL, Filipczyk A & Pera MF (2003). Characterization and Culture of Human Embryonic Stem Cells. Trends In Cardiovascular Medicine 13(7):295-301.
  2. Martin F. Pera, Adam Filipczyk, Susan M. Hawes, and Andrew L. Laslett (2003) Isolation, characterization, and differentiation of human embryonic stem cells. Methods in Enzymology 365: 429-46.
  3. Martin F. Pera, Andrew Laslett, Susan M. Hawes, Irene Tellis, Karen Koh, and Linh Nguyen. (2006) Isolation and characterization of human embryonic stem cells.  In Embryonic stem cells:  a practical approach.  Edited by Elena Notarianni and Martin J. Evans. Oxford University Press, p238-259.
  4. Herszfeld D, Wolvetang E, Filipczyk A, Langton –Bunker E, Koh K, Nguyen L, Tellis I, Michalska A, Laslett AL, Reubinoff BE, Houssami S, Looijenga LH and Pera MF (2006) CD30 is a survival factor and a biomarker for transformed human pluripotent stem cells. Nature Biotechnology 24(3):351-7.  
  5. Andrew L. Laslett, Sean Grimmond, Brooke Gardiner, Lincon Stamp, Adelia Lin, Susan M. Hawes, Sam Wormald, David Nikolic-Paterson, David Haylock, and Martin F. Pera (2007)  Transcriptional Analysis of Early Lineage Commitment In Human Embryonic Stem Cells.  BMC Developmental Biology Mar 2;7:12.
  6. Adam A. Filipczyk, Andrew L. Laslett, Christine Mummery & Martin F. Pera. (2007) Differentiation is Coupled to Changes in the Cell Cycle Regulatory Apparatus of Human Embryonic Stem Cells. Stem Cell Research 1:1; 45-60.
  7. Andrew L. Laslett, Adelia Lin and Martin F. Pera (2007) Characterization and differentiation of human embryonic stem cells. In: Embryonic Stem Cells edited by .JR.Masters et al, Springer, p27–40.
  8. Andrew L. Laslett, Andrew Fryga and Martin F. Pera (2007) Flow cytometric analysis of human embryonic stem cells. In: Human Stem Cell Manual:  A Laboratory Guide, edited by Jeanne F. Loring, Elsevier, p96-107.
  9. Lina Gubhaju, Andrew Laslett, John F. Bertram, Anthony Zulli, Jane M Black (2008) Immunohistochemical localisation of TRA-1-60, TRA-1-81, GCTM-2 and podocalyxin in the developing baboon kidney. Histochemistry and Cell Biology 129(5):651-7.
  10. Kolle, G. Ho, SHM., Zhou, Q, Chy, H.S., Krishnan, K, Cloonan, N, Bertoncello, I, Laslett, A.L. and Grimmond, S.M.  Identification of human embryonic stem cell surface markers by combined membrane-polysome translation state array analysis and immunotranscriptional profiling. Stem Cells (in Press, July 2009)