Use of the bnx/hu xenograft model of human hematopoiesis to optimize methods for retroviral-mediated stem cell transduction (Review).

M. A. Dao, Jan Nolta

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

The potentiality of primitive human hematopoietic cells can be profoundly affected by in vitro culture. Due to the growing number of protocols proposed for stem cell gene therapy and ex vivo expansion, it is crucial to define methods to preserve the generative capacity of human stem cells in culture while promoting self-renewal divisions. Stem cell division, homing, and subsequent lineage development can only be studied definitively by marking of pluripotent cells, followed by tracking and clonal analysis of the progeny in a long-term transplantation system. We have developed a bnx/hu xenograft model, in which transduced human hematopoietic cells can be individually tracked into different lineages over the course of one year post-transplantation. The tracking is accomplished by single cell cloning of individual T lymphoid and myeloid progenitors recovered from the marrow of the mice, and clonal integration analysis by the sensitive technique of single-colony inverse PCR. All cells derived from a stem cell transduced by a retroviral vector will carry the unique restriction fragment length polymorphism (RFLP) created by the random integration event. We have used the bnx/hu xenograft system coupled with single-colony inverse PCR to determine that human stem cells require stromal support, fibronectin support with cytokines, or the presence of Flt3 ligand during a 72-h ex vivo culture to maintain the ability to sustain long-term multilineage hematopoiesis.

Original languageEnglish (US)
Pages (from-to)257-264
Number of pages8
JournalInternational Journal of Molecular Medicine
Volume1
Issue number1
StatePublished - Jan 1998
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

Fingerprint Dive into the research topics of 'Use of the bnx/hu xenograft model of human hematopoiesis to optimize methods for retroviral-mediated stem cell transduction (Review).'. Together they form a unique fingerprint.

  • Cite this