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First published online October 26, 2006
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2005-0639v1
25/2/529    most recent
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Submitted on December 20, 2005
Accepted on October 18, 2006

Technology Development

ENHANCED IN VIVO HOMING OF UNCULTURED AND SELECTIVELY AMPLIFIED CORD BLOOD CD34+ CELLS BY CO-TRANSPLANTATION WITH CORD BLOOD DERIVED UNRESTRICTED SOMATIC STEM CELLS

Shing Leng Chan 1, Michael Choi 1, Stephan Wnendt 2, Morey Kraus 2, Eileen Teng 1, Hwei Fen Leong 1, Shoshana Merchav 1*

1 ViaCell Singapore Research Centre, Singapore, Singapore
2 ViaCell Inc, Cambridge, Massachusetts

* To whom correspondence should be addressed. E-mail: Smerchav{at}viacellinc.com.


  Abstract

Mesenchymal stem cells have been implicated as playing an important role in stem cell engraftment. Recently a new pluripotent population of umbilical cord blood (UCB) cells, Unrestricted Somatic Stem Cells (USSCs), with intrinsic and directable potential to develop into mesodermal, endodermal and ectodermal fates, has been identified. In this study we evaluated the capacity of ex-vivo expanded USSCs to influence the homing of UCB-derived CD34+ cells into the marrow and spleen of NOD/SCID mice.

USSCs induced a significant enhancement of CD34+ cell homing to both bone marrow and spleen (2.2 ± 0.3 and 2.4 ± 0.6 -fold, respectively; p<0.05), with a similar magnitude induced by USSCs that had been thawed prior to transplantation. The effect of USSCs was dose-dependent and detectable at USSC/CD34+ ratios of 1:1 and above. Enhanced marrow homing by USSCs was unaltered by extensive culture passaging of the cells, as similar enhancement was observed for both early (p5) and late (p10) passage USSCs.

The homing effect of USSCs was also reflected in an increased proportion of NOD/SCID mice exhibiting significant human cell engraftment six weeks following transplantation, with a similar distribution of myeloid and lymphoid components.

USSCs enhanced the homing of cellular products of ex-vivo expanded UCB lineage-negative (lin-) cells, generated in 14-day cultures by Selective Amplification. The relative proportion of homing CD34+ cells within the culture-expanded cell population, was unaltered by USSC co-transplantation.

Production of Stromal Derived Factor -1 (SDF-1) by USSCs was detected by both gene expression as well as protein released into culture media of these cells. Knockdown of SDF-1 production by USSCs using lentiviral-SiRNA, led to a significant (P<0.05) reduction in USSC-mediated enhancement of CD34+ homing.

Our findings thus suggest a clinical potential for using USSCs in facilitating homing and engraftment for all cord blood transplant recipients.

 Key Words. USSC, enhanced homing, CD34+ cells, SDF-1 knockdown




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