Module 2: AIBN Bioreactor Program

Module member at the University of Queensland

Module Leaders Professor Peter Gray

Professor Justin Cooper-White

Associate Professor Ernst Wolvetang
Host Organisation
 Australian Institute for Biotechnology and Nanotechnology, Univerity of Queensland

Module description

The overall aim of this module is to develop defined, animal product free and scaleable bioprocesses for the expansion of human embryonic stem cells and subsequent differentiation of these cells.

The research team is multidisciplinary, with expertise in stem cell biology (Wolvetang), biomaterials and microfabrication (Cooper-White) and mammalian cell culture and bioprocess development (Gray). The module continues the development of novel products and bioprocesses with application to researchers working in the stem cell field, and to researchers developing potential therapeutics based on stem cell technology.

Aims

  1. Generation of lineage specific reporter lines to allow high throughput analysis of lineage specific differentiation in hESC and EBs in situ.
  2. Development of defined component biomimetic surfaces to replace the current complex animal derived products such as Matrigel used currently  for the feeder free cultivation of hESC.
  3. Translation of the optimal biomimetic surfaces from aim 2 into three dimensional bead (solid and porous) and porous scaffold formats. Importantly, these biomimetic surfaces will be composed of or incorporate segments to allow controlled release of hESCs at desired time points for further use in aim 5 and 6.
  4. Optimisation by factorial design of soluble maintenance factors that support undifferentiated expansion of hESC in defined media and in the absence of feeder cells.
  5. Development of diagnostic microbioreactors for the rapid screening and assessment of hESC growth on and in the presence of a range of soluble and bound factors and under differing microenvironmental conditions in order to determine optimal process conditions required for aim 6.  
  6. Development of a bioprocess based on information obtained in aims 1 to 5 above which is capable of 108 fold expansion of hESC under fully defined, animal protein free conditions whilst maintaining pluripotency as indicated by in-vivo and in-vitro markers. Use of genomics and metabolomics to accurately describe the state of hESC when they are growing under conditions which maintain pluripotency.
  7. Development of a process capable of handling the formation and subsequent cultivation of 100,000 hESC embryoid bodies suitable for optimisation of haematopoietic differentiation developed in the Elefanty/Stanley lab, and other lineages including,  osteogenic, chondrogenic, myogenic and neurogenic differentiation, utilising the lineage specific reporter lines developed in aim 1.

Module Leader biographies

Professor Cooper-White’s research interests are in biomaterials synthesis and processing, surface engineering, tissue engineering, non-Newtonian fluid mechanics, microfluidics and microbioreactors. He is currently a group leader of the Australian Institute for Bioengineering and Nanotechnology at the University of Queensland and manages a group of 6 postdoctoral fellows and 14 students. He has authored or co-authored over 190 research publications and presentations, including over 90 refereed publications and is often asked to present plenary, keynote and invited lectures at national and international conferences.

Professor Cooper-White is a consultant for a number of international companies, Associate Editor of the Korean-Australian Rheology Journal, on the Editorial Boards of Rheologica Acta, Soft Materials  and the Open Biomedical Engineering Journal. He holds seven patents in the areas of formulation design for agriproducts, microbioreactors, particle synthesis using microfluidic devices and tissue engineering scaffolds. He has held the position of  President of both the Australasian Society for Biomaterials and Tissue Engineering  (2006-2008) and of the Australian Society of Rheology (2002-2004).

Professor Peter Gray was appointed in 2003 as the inaugural Director of the Australian Institute of Bioengineering and Nanotechnology (AIBN) at the University of Queensland. Professor Gray is also Professor of Biotechnology at the University of New South Wales, and was Director of the Bioengineering Centre, UNSW and Senior Principal Research Fellow at the Garvan Institute of Medical Research. He has held academic positions at University College London and at the University of California, Berkeley. Professor Gray has had commercial experience in the USA working for Eli Lilly and Co and the Cetus Corporation.

Professor Gray serves on the Boards of Biopharmaceuticals Australia Pty Ltd, ACYTE Biotechnology Pty Ltd and the Advanced Water Management Centre, and on a number of government committees in the fields of biotechnology, pharmaceuticals and education. He is a regular reviewer and consultant for public and private sector research initiatives in Australia and overseas.

Associate Professor Ernst Wolvetang leads the human embryonic stem cell engineering group within the Australian Institute for Bioengineering and Nanotechnology at the University of Queensland. Previously, he was head of the Basic Human Embryonic Stem Cell Biology Laboratory at the Australian Stem Cell Centre. Between 1997 and 2006, Associate Professor Wolvetang was located at the Centre for Early Human Development in the Institute for Reproduction and Development, Monash University.

He then joined the laboratory of Associate Professor Martin Pera (2003-2006) to investigate the role of signalling/gene regulation pathways in the control of growth, differentiation and apoptosis of human embryonic stem cells (hESC) by using retroviral transduction of sh RNA’s and cDNAs. This work culminated in his publication (Nature Biotechnology) describing the relationship between the expression of CD30, apoptosis and genetic instability in hESC. As a result of this work, Associate Professor Wolvetang is now recognised nationally and internationally for his knowledge on proliferation, apoptosis and genetic stability of hESC.

Contact details

Professor Peter Gray     
 E-mail  peter.gray@uq.edu.au
 Phone  +61 7 3346 3888

Professor Justin Cooper-White
 E-mail  j.cooperwhite@uq.edu.au
 Phone   +61 7 3346 3858

Associate Professor Ernst Wolvetang
E-mail 
 e.wolvetang@uq.edu.au
Phone  +61 7 3346 3894

Selected publications

  1. Vari, F, Munster, DJ, Hsu, JL, Rossetti, TR, Mahler, SM, Gray, PP, Turtle, CJ, Prue, RL and Hart DNJ (2008): “Practical blood dendritic cell vaccination for immunotherapy of multiple myeloma”. British Journal of Haematology. 143 (3), 374-377. IF=3.73 Citations: 0
  2. Xu ZP, Niebert M, Porazik K, Walker TL, Cooper HM, Middelberg APJ, Gray PP, Bartlett PF, Lu GQM (2008): "Subcellular Compartment Targeting of Layered Double Hydroxide Nanoparticles". J Control Release. 130(1), 86-94. IF= 5.2 Citations: 1
  3. Xu ZP,Stevenson GS, Lu CQ, Bartlett PF, Gray PP (2006) Stable suspension of layered double hydroxide nanoparticles in aqueous solution. J. Am Chem Soc.128(1), 36-7. IF=7.9   Citations: 36
  4. Prowse ABJ, McQuade LR, Bryant KJ, Marcal H, Gray PP(2007). Identification of Potential Pluripotency Determinants for Embryonic Stem Cells Following Proteomic Analysis. Journal of Proteome Research, 6 (9), 3796- 3807. IF= 6.8 Citations: 8
  5. Croll, Tristan I.,  O'Connor, Andrea J.,  Stevens, Geoffrey W.,  Cooper-White, Justin J.,  ‘A blank slate? Layer-by-layer deposition of hyaluronic acid and chitosan onto various surfaces’, Biomacromolecules, 7(5), 1610-1622, 2006..
  6. Rowlands Andrew S; George Peter A; Cooper-White Justin J.,  ‘Directing osteogenic and myogenic differentiation of MSCs: interplay of stiffness and adhesive ligand presentation.’    American journal of physiology. Cell physiology   295(4),  C1037-44 2008.
  7. George P.A., Donose B.C., Cooper-White J.J., Self-assembling polystyrene-block-poly(ethylene oxide) copolymer surface coatings: Resistance to protein and cell adhesion BIOMATERIALS    30 (13)  2449-2456   2009.
  8. Wolvetang E ,Herszfeld D, Langton –Bunker E, Chung T, Filipczyk A, Houssami S, Koh K, Laslett AL, Michalska A, Nguyen L, Reubinoff BE, Tellis I, Auerbach JM, Ording CJ, Looijenga LHJ, and Pera MF. (2006) CD30 is a survival factor and a biomarker for transformed human pluripotent stem cells. Nat.Biotech. 24(3):351-7.
  9. Grandela C and Wolvetang E. (2007) hESC Adaptation, Selection and Stability. Stem Cell Rev. 2007 Fall;3(3):183-91.
  10. Grandela C, Pera MF and Wolvetang EJ. p53 is required for etoposide-induced apoptosis of human embryonic stem cells. (2007) Stem Cell Research 1 (2) Pages 116-128.
  11. Hannan, NRF and EJ Wolvetang. Adipocyte differentiation in human embryonic stem cells transduced with Oct4 shRNA lentivirus. (2008) Stem Cell Dev. Sep 2.
  12. A B J Prowse, E J Wolvetang, P P Gray. A rapid, cost effective method for counting human embryonic stem cell numbers as clumps. (2009) Biotechniques (in press) [IF 2.759 Cited 0].
  13. A B J Prowse, J Wilson, G Osborne, P P Gray, EJ Wolvetang. Multiplexed staining of live human embryonic stem cells for flow cytometric analysis of Pluripotency markers. (2009) Stem Cells and Development (in press) [IF 3.48; Cited 0].
  14. N Hannan, P Jamshidi, MF Pera, EJ Wolvetang. BMP -11 and Myostatin Maintain Human Embryonic Stem Cells in Feeder Free Cultures. (2009) Stem Cells and cloning [2.937 Cited 0 ] (In Press)