Purified recombinant TAT-homeobox B4 expands CD34(+) umbilical cord blood and peripheral blood progenitor cells ex vivo.
Hematopoietic stem cells (HSCs) are commonly used in clinical transplantation protocols to treat a variety of diseases. However, efficient transplantation requires a substantial amount of HSCs from different sources and may require expansion. Therefore, effective expansion of HSCs remains a technical hurdle blocking the development of advanced cell therapies. The product of the human homeobox B4 (HOXB4) gene was recently demonstrated to effectively expand HSCs from umbilical cord blood (UCB) or bone marrow in either a retroviral or recombinant protein form. Our study purified TAT-HOXB4 proteins and demonstrated their ability to expand UCB and peripheral blood (PB) progenitor cells. The results showed that the TAT-HOXB4 gene product expanded the CD34(+) progenitor cells from UCB and PB by approximately 7.5-fold. The results from a semisolid cloning assay, a human long-term culture-initiating cell assay, and a nonobese diabetic-severe combined immunodeficiency mice repopulating assay showed that TAT-HOXB4 expanded hematopoietic progenitor cells while retaining their repopulating capacity and multipotency. TAT-HOXB4 protein also expanded engrafted stem cells that were previously expanded in a secondary transplantation assay. The results demonstrated the feasibility of using TAT-HOXB4 to expand UCB and PB progenitor cells, which are readily available to treat different hematological malignancies and nonhematological diseases.
Huang
Chi-Hung
CH
Institute of Biochemistry, National Chung-Hsing University, Taichung, Taiwan.
Chen
Po-Min
PM
Lu
Tsung-Chi
TC
Kung
Wen-Mei
WM
Chiou
Tzeon-Jye
TJ
Yang
Muh-Hwa
MH
Kao
Jung-Yie
JY
Wu
Kou-Juey
KJ
D000818:Animals; D018952:Antigens, CD34 / Q000276:immunology*; D016895:Culture Media, Serum-Free; D024202:Electrophoretic Mobility Shift Assay; D005312:Fetal Blood / Q000276:immunology*; D005434:Flow Cytometry; D015756:Genes, tat; D018398:Homeodomain Proteins; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D016133:Polymerase Chain Reaction; D013234:Stem Cells / Q000276:immunology*
D018952:Antigens, CD34; D016895:Culture Media, Serum-Free; D018398:Homeodomain Proteins