Module 3: Functional Assessment of ex vivo Expanded CB HSC and HPC

Human and murine platelets in PB of NOD/SCID mice

Module Leader Associate Professor Susie Nilsson
Host Organisation CSIRO and CRC-Polymers

Module description

This module is divided into two components, which involve the in vivo functional assessment of haemopoietic stem cells (HSC) expanded in smart surface bioreactors and the ability of HSC whose differentiation has been directed to committed megakaryocytic progenitors to abrogate thrombocytopenia in vivo. Initially, work with HSC expansion will involve populations of CB CD34+ cells grown on surfaces created by Professor Justin Cooper-White at the Australian Institute for Nanotechnology and Biotechnology. Functional studies will be performed using well validated approaches currently used for assessing adult HSC and will involve a comparison with known populations of cord blood (CB) and adult mobilised peripheral blood (mPB) stem and progenitor cells. Work involving megakaryocytic progenitors will assess donor derived platelet production in murine models of human haemopoiesis.

These studies are essential prerequisites for cells to be considered for clinical cell transplantation.

Bone marrow multilineage reconstitution following CB HSC transplant into NOD/SCID mice

A whole bone marrow with WBC gate (R1), B CD45 expression of R1, C multilineage analysis of R1

Aims

This module is designed to test the hypothesis that ex vivo expanded hemopoietic stem cell (HSC) will be functionally equivalent to their counterparts prospectively isolated from CB and mPB and that differentiated megakaryocyte precursors will decrease the severity of thrombocytopenia when tested in in vivo murine models of human hemopoiesis.

This hypothesis will be tested using the following two specific aims:

  1. to determine the hemopoietic potential of HSC expanded ex vivo on different smart surfaces
  2. to determine whether megakaryocytic (Mk) progenitor cells differentiated in a two phase culture system home to the bone marrow and give rise to sustained platelet production post-transplant.
      

Module Leader biography

In July 2009 Associate Professor Susie Nilsson (nee Begg) joined CSIRO Molecular and Health Technologies but along with her team remains based in the ASCC laboratories. Prior to this, Associate Professor Nilsson was the head of the Niche Laboratory at the Australian Stem Cell Centre having moved from the Peter MacCallum Cancer Centre at the end of 2005, where she was head of the Microenvironment Laboratory within the Stem Cell Program.

Associate Professor Nilsson’s underpinning scientific objective has been to characterise the haemopoietic stem cell niche. She developed an in vivo cell tracking model to identify haemopoietic stem cells (HSC) in situ and was one of the first investigators to provide convincing evidence that HSC preferentially seek and reside within the endosteal region of the bone marrow.

Associate Professor Nilsson is the author of 45 publications, including 32 in the past 10 years. During this time she has been invited to submit five peer reviewed papers and five book chapters.

In the past decade, she been awarded six patents, all of which are at various stages from provisional applications to national phase. She is currently a member of the Board of Directors of Experimental Haematology and a member of the International Society for Stem Cell Research.

Contact details

 E-mail  susie.nilsson@csiro.au
 Phone  +61 3 9271 1150

Selected publications

  1. S.K. NILSSON, D.N. Haylock, H.M. Johnston, T. Occhiodoro, T.J. Brown, P.J. Simmons. (2003) Hyaluronan is synthesised by primitive hemopoietic cells, participates in their lodgement at the endosteum following transplantation and is involved in the regulation of their proliferation and differentiation in vitro. Blood 101:856-62.
  2. R.L. Driessen, H.M. Johnston, S.K. NILSSON. (2003) Membrane-Bound Stem cell Factor Is a Key Regulator in the Initial Lodgement of Stem Cells within the Endosteal Marrow Region. Exp Hematol 12: 1284-1291.
  3. S.K. NILSSON, H.M. Johnston, G.A. Whitty, B. Williams, R.J. Webb, D.T. Denhardt, I. Bertoncello, L.J. Bendall, P.J. Simmons, D.N. Haylock (2005) Osteopontin, a Key Component of the Hematopoietic Stem Cell Niche and Negative Regulator of Primitive Hematopoietic Progenitor Cells. Blood 106:1232-1239.
  4. D.N. Haylock, S.K. NILSSON (2005) Stem cell regulation by the haemopoietic stem cell niche. Cell Cycle 4:1353-1355.
  5. D.N. Haylock, S.K. NILSSON (2006) Osteopontin: A bridge between bone and blood. Br. J. Haematol. 134:467-74.
  6. D.N. Haylock, S.K. NILSSON (2006) The role of hyaluronic acid in hemopoietic stem cell biology. Reg. Med. 1:437-445.
  7. D.N. Haylock, B. Williams, H.M. Johnston, M.C.P Liu, K.E. Rutherford, G.A. Whitty, P.J. Simmons, I. Bertoncello, S.K. NILSSON. (2007) HSC with higher hemopoietic potential reside at the bone marrow endosteum. Stem Cells. 25:1062-1069.
  8. S.K. NILSSON, H.M. Prince, D. Wall, D.N. Haylock. (2007) Recent Australian Experience with Haemopoietic Stem and Progenitor Cell Expansion. Cytotherapy. 9:231-235.
  9. D.N. Haylock, S.K. NILSSON (2007) Expansion of umbilical cord blood for clinical transplantation. Current Stem Cell Research & Therapy. 2:324-335.
  10. J. Grassinger, D.N. Haylock, M. Storan, G.O. Haines, B. Williams, G.A. Whitty, A. Vinson, C.L. Be, S. Li, E.S. Sørensen, P.L. Tam, D.T. Denhardt, D. Sheppard, P.F. Choong, S.K. NILSSON (2009) Osteopontin is critical for the attraction, retention, regulation and release of hemopoietic stem and progenitor cells to, in and from their bone marrow niche in a process mediated by α9β1 and α4β1 integrins. Blood. 114:49-59.