Project description:Efficient bone marrow homing is a prerequisite for successful engraftment of transplanted HSPC. This study aims at determining factors important in homing of these cells from the blood to the marrow and their re-engraftment. We have isolated nucleated cells from mobilized peripheral blood stem cell harvests (PBSC). CD34+ hematopoietic stem and progenitor cells (HSPC) were enriched from PBSC harvests by immunomagnetic separation using negative selection method. Enriched cells were cultured in vitro for expansion in presence or absence of cytokine (GF: SCF+TPO+FLT3L) and/or chemokine (homing factor SDF1) mixture for 8 days at 370C in humidified atmosphere of 5% CO2 in air. GF+SDF1 –stimulated HSPC showed significantly increased total nucleated ells as well as CD34+ cells as compared to GF-stimulated HSPC as well s unstimulated cultured HSPC. On transplantation of these cells in vivo in mice model who were previously irradiated at sublethal dose of 375cGy, it was observed that GF+SDF1-stimulated HSPC exhibited significantly better engraftment in terms of presence of human CD45+ cells in mouse blood at 6 week post –transplantation. We have compared gene expression profiles of mobilized PBSC harvest cells, enriched CD34+ HSPC population and HSPC cultured in plain media, in presence with GF and in presence of GF+SDF1 derived from mobilized PBSC harvests with respect to genes involved in homing and engraftment capacities.
Project description:A prospective randomized trial has shown that there is a survival advantage for allogeneic transplant patients receiving Granulocyte Colony Stimulating Factor (G-CSF) stimulated peripheral blood mononuclear cells (GPBMC) versus bone marrow (BM) as a source of stem cells. The biological basis for this advantage is not clear, and may be attributable to qualitative as well as quantitative differences in the CD34 cells, T-cells and/or the monocytes transplanted. To begin to address this issue, gene expression patterns in monocytes isolated from G-CSF mobilized peripheral blood were compared those from normal, non-mobilized peripheral blood to identify functional pathways that may distinguish these two populations. Keywords: Cell type comparison
Project description:Human naive T cells from peripheral blood were cultured in 24 wells coated with anti-CD3 and anti-CD28 antibodies in the presence or absence of retinoid acid, IL-12, and 1,25 (OH)2 vitamin D3. The T cells were FACS-sorted based on expression of CD3, integrin alpha4beta7, cutaneous lymphocyte antigen (CLA) and chemokine receptor 10. This serie includes microarray data from stimulated T cells under indicated conditions. Keywords: Human T cell, vitamin D and A, chemokine receptor, nuclear receptor
Project description:Investigation of human hematopoietic stem cells gene expression patterns originating from different stages of ontogeny including fetal blood, cord blood, bone marrow, and mobilized peripheral blood in Lin-CD34+CD38- versus Lin-CD34+CD38+ populations. Keywords: other
