Stem Cell Laboratory

Group Leaders Dr Mirella Dottori, Principal Investigator Dr Alice Pébay, Principal Investigator Postdoctoral Fellows Dr Mark Denham Dr Brock Conley Research Assistant Ms Jessie Leung Student Ms Cheryl Tay, PhD student (Monash University)
	Stem Cell Laboratory Staff (L/R Alice Pébay, Brock Conley, Mirella Dottori and Jessie Leung [Mark Denham not shown in photo])

Major Laboratory Interests

• Involvement of lysophospholipids in stem cell biology
• Differentiation of neural stem cells derived from human embryonic stem cells

Brief Description of Projects

Involvement of lysophospholipids in stem cell biology:

Lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are bioactive lysophospholipids that act on a wide range of cells and regulate numerous cellular functions. They have been clearly identified to have widespread physiological and pathophysiological actions, controlling events within the nervous, reproductive, gastrointestinal, vascular, respiratory and immune systems, and also having a prominent role in cancer, early mammalian embryogenesis and stem cells. LPA and S1P are also molecules of inflammation. As they are synthetised by platelets and released upon activation, their concentrations increase during inflammation and trauma. In the nervous system, their levels increase in pathological conditions where the blood brain barrier integrity is damaged, making them significant factors contributing to an inflammatory response during neurotrauma. This project focuses on the understanding of the effects of LPA and S1P in stem cells, neural stem cell (NSC) and their derivatives and aims at identifying the signaling pathways activated. Indeed, modulating LPA and S1P signallings may have a significant impact in injury within the central nervous system, allowing new avenues for potential therapeutic approaches.

Differentiation of neural stem cells derived from hESC:

Neural stem cells (NSC) can be easily derived from adult and fetal tissue, as well as directly differentiated from embryonic stem cells. In the human, the signalling factors and mechanisms that regulate NSC maintenance and their further differentiation into neural and glial progenitor subtypes still remain largely unknown. The use of hESC neural derivatives may provide important breakthroughs in medical research by providing cultured human cells that can be used as in vitro models to study neurodegenerative disorders. If hESC are to be used to study of early human neural development and disease, it is essential to understand the key pathways involved that regulate hESC neuronal differentiation. In our laboratory we have established an in vitro model of human NSC differentiation. Using a range of culture systems, we can monitor the various stages of neuronal differentiation from hESC to mature neurons and glia, which is very useful for exploring the signals involved in regulating this process. Our major aims are to direct differentiation of hESC towards specific neural lineages, including neural crest derivatives, dopaminergic neurons and oligodendrocytes, and to identify key factors that regulate this process. From these studies we can regulate hESC neural differentiation and derive specific neural cell types to be used for further research and clinical studies.

Techniques Used

• Cell culture
• Neural differentiation assays
• Proliferation assays
• Apoptosis and necrosis assays
• Migration assays
• ELISA
• Immunostaining
• Western-blots
• FACS analysis
• RT-PCR
• Transfection
• Lentiviral transduction

Recent Publications

Hotta R, Pepdjonovic L, Anderson RB, Zhang D, Bergner AJ, Leung J, Pébay A, Young HM, Newgreen DF, Dottori M. (2009) Small Molecule Induction of Neural Crest-like Cells derived from Human Neural Progenitors. Stem Cells In press, accepted 16-8-09

Wong R, Donovan PJ and Pébay A (2009). Molecular mechanism involved in the maintenance of pluripotent stem cells; Pluripotent Stem Cells. F. Colombus, Editor. Nova Publishers. In press, accepted 29.07.09

Pitson S and Pébay A (2009). Regulation of stem cell pluripotency and neural differentiation by lysophospholipids. Neurosignals,17(4).

Pébay A and Pera MF (2009). Growth Factors and the Serum-free Culture of Human Pluripotent Stem Cells. Essentials of Stem Cell Biology, Second Edition. R. Lanza, Editor. Elsevier Academic Press. In press, accepted 08.01.09

Denham, M. and Dottori, M. (2009) Signals Involved in Neural Differentiation of Human Embryonic Stem Cells. NeuroSignals In press, accepted 18-6-09.

Davidson KC, Wong R, Leung J, Pera MF and Pébay A (2009). Acute effect of endothelins on intercellular communication of human embryonic stem cells. Journal of Stem Cells 4(1).

Wong R and Pébay A (2009). Study of gap junctions in human embryonic stem cells. Methods in Molecular Biology; Human Embryonic Stem Cells: Methods and Protocols, second edition. Chapter 12, 211-228. K. Turksen, Editor. Humana Press.

Dottori M, Pébay A and Pera MF (2009). Neural Differentiation of Human Embryonic Stem Cells. Methods in Molecular Biology; Protocols for Neural Cell Culture: Fourth Edition. Chapter 4, 75-86. L. Doering, Editor. Humana Press.

Niclis JC, Trounson AO, Dottori M, Ellisdon AM, Bottomley SP, Verlinsky Y, Cram DS. (2009) Human Embryonic Stem Cell Models of Huntington’s Disease, Reproductive Biomedicine Online 19, 106-113.

Bertram C, Hawes SM, Egli S, Peh GSL, Dottori M, Kees UR and Dallas PB. (2009) Effective adenovirus mediated gene transfer into neural stem cells derived from human embryonic stem cells. Stem Cells and Development In press, accepted 13-7-09.

Dottori M, Leung J, Turnley AM and Pébay A (2008). Lysophosphatidic acid inhibits neuronal differentiation of neural stem/progenitor cells derived from human embryonic stem cells. Stem Cells 26 (5),1146-1154.

Dottori M and Pera M.F. (2008) Neural differentiation of human embryonic stem cells. In: Methods in Molecular Biology, Neural Stem Cells, Second Edition. Vol 438, 19-30.  L.P.Weiner, Editor. The Humana Press Inc.

Wong R, Pera M and Pébay A (2008). Role of gap junctions in embryonic and somatic stem cells. Stem Cell Reviews, 4(4), 283-292.

Davidson K, Dottori M and Pébay A (2008). Human embryonic stem cells: key characteristics and main applications in disease research. Chapter 3, 155-187. Cell Applications in Diseases. M. L. Sorensen Editor. Nova Science Publishers.

Wong R and Pébay A (2008). Signaling pathways involved in the maintenance of human embryonic stem cells. Stem Cell Research Compedium, Volume 1.Chapter 19, in press. P. Koka, Editor. Nova Publishers.

Wong R and Pébay A (2008). Signaling pathways involved in the maintenance of human embryonic stem cells. Leading-Edge Stem Cell Research.Chapter 8, 123-138. P. Koka, Editor. Nova Publishers.

Pébay A, Bonder C and Pitson S (2007). Stem cell regulation by lysophospholipids. Prostaglandins & other lipid mediators, 84(3-4), 83-97.

Wong R, Tellis I, Jamshidi P, Pera M and Pébay A (2007). Anti-apoptotic effect of sphingosine-1-phosphate and platelet-derived growth factor in human embryonic stem cells. Stem Cells and Development, 16(6). 989-1002.

Costa M*, Dottori M*, Sourris K, Jamshidi P, Hatzistavrou T, Davis R, Azzola L, Jackson S, Lim SM, Pera MF, Elefanty AG and Stanley EG. A Method for genetic modification of human embryonic stem cells using electroporation. (2007) Nature Protocols  2, 792-796. *Equal first author.

Davidson KC, Jamshidi P, Daly R, Hearn MT, Pera MF and Dottori M. Wnt3a regulates survival, expansion, and maintenance of neural progenitors derived from human embryonic stem cells. (2007) Mol Cell Neuro 36, 408-415.

Wong R and Pébay A (2006). Signaling pathways involved in the maintenance of human embryonic stem cells. Journal of Stem Cells 1 (4), 271-282.

Wong R, Dottori M, Koh K, Nguyen L, Pera M and Pébay A (2006). Gap junctions modulate apoptosis and colony growth of human embryonic stem cells maintained in a serum-free system. Biochemical and Biophysical Research Communications 344, 181-188.

Rouach N, Pébay A, Meme W, Cordier J, Ezan P, Etienne E, Giaume C and Tence M. (2006). S1P inhibits gap junctions in astrocytes: involvement of G and Rho GTPase/ROCK. European Journal of Neuroscience 23(6), 1453-1464.

Trounson A and Pébay A (2006). A Role for Neurotrophins in Embryonic Stem Cell Growth. Commentary Developmental Cell 10, 158-159.

Pébay A and Pera M (2006). Growth factors and the serum-free culture of human pluripotent stem cells. Essentials of Stem Cell Biology, Part four, Chapter 41, 313-316.  R. Lanza, Editor. Elsevier Academic Press.

Pébay A, Wong R, Pitson S, Wolvetang E, Peh G, Filipczyk A, Koh K, Tellis I, Nguyen L and Pera M. (2005). Essential roles of sphingosine-1-phosphate and platelet-derived growth factor in the maintenance of human embryonic stem cells. Stem Cells 23 (10), 1541-1548.

Denham M, Huynh T, Dottori M, Allen G, Trounson A and Mollard R. Neural stem cells express non-neural markers during embryoid body co-culture. (2005) Stem Cells 24, 918-927.

Costa M*, Dottori M*, Ng E, Hawes SM, Sourris K, Jamshidi P, Pera MF, Elefanty AG and Stanley EG. The hESC line Envy expresses high levels of GFP in all differentiated progeny. (2005) Nature Methods 2, 259-260. (*Equal first author)

Pera M and Dottori M. Stem cells and their development potential. (2005) In: Stem Cells From Bench to Bedside. Eds A. Bongso and E.H Lee. World Scientific Press.

Wong R, Pébay A, Koh K, Nguyen L and Pera M. (2004). Presence of Functional Gap Junctions in Human Embryonic Stem Cells. Stem Cells 22 (6), 883-889.

Contact us

Stem Cell Laboratory
Centre for Neuroscience

apebay@unimelb.edu.au (Dr Alice Pebay)

mdottori@unimelb.edu.au (Dr Mirella Dottori)

http://www.cns.unimelb.edu.au (The Centre for Neuroscience)

http://www.pharmacology.unimelb.edu.au (Department of Pharmacology)