Isolation and tracking of a rare lymphoid progenitor cell which facilitates bone marrow transplantation in mice.
Bone marrow cells are composed of pluripotent stem cells to terminally differentiated cells, with a wide variety of abundance of each cell type. In the past, many of the cell types within this heterogeneous population have been characterized either by expression of specific proteins or using functional markers. In spite of promising results obtained with the latter method, various cell types within bone marrow have not been well characterized due to the low abundance of a specific cell type. Considering the demand for a reliable technique to enrich cell types, a wide variety of approaches, ranging from simple nylon wool columns to high-speed cell sorting, have evolved. Only limited success has been obtained with approaches ranging from the detection of MHC antigen to positron emission tomography to track the ontogeny of specific bone marrow-derived cells in studies of syngeneic or allogeneic transplantation. The present study describes a relatively simple method to enrich and track a rare bone marrow cell (facilitating cell, FC), which can facilitate allogeneic bone marrow stem cell transplantation in mice. The isolation technique is comprised of enrichment of FC by magnetic activated cell sorting (MACS) system followed by purification through high-speed cell sorter. An initial inoculation of 30,000 FC obtained from male mice was detected in the thymus, spleen, and bone marrow of allogeneic female recipients, by using 32P-labeled dCTP in a specific PCR for Y-chromosome. This technique may improve the efficiency of isolation of other rare cells from the bone marrow.
Gangopadhyay
Nupur N
NN
Division of Thoracic Surgery, Department of Surgery, School of Medicine, University of Pittsburgh, W1554 BST, 200 Lothrop Street, Pittsburgh, PA 15213, USA. gangopadhyayn@msx.upmc.edu
Shen
Hongmei
H
Landreneau
Rodney
R
Luketich
James D
JD
Schuchert
Matthew J
MJ
D000818:Animals; D001854:Bone Marrow Cells / Q000502:physiology*; D016026:Bone Marrow Transplantation; D018414:CD8-Positive T-Lymphocytes / Q000502:physiology*; D005260:Female; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D018189:Immunomagnetic Separation / Q000379:methods*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D011948:Receptors, Antigen, T-Cell / Q000032:analysis; D014184:Transplantation, Homologous
D011948:Receptors, Antigen, T-Cell