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Developmental Immunology
Volume 4 (1995), Issue 4, Pages 235-246

Enrichment of Early Fetal-Liver Hemopoietic Stem Cells of the Rat Using Monoclonal Antibodies Against the Transferrin Receptor, Thy-1, and MRC-Oχ82

1Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, United Kingdom
2MRC Human Genetics unit, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, Scotland

Received 14 December 1994; Accepted 28 June 1995

Copyright © 1995 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Fetal livers from inbred rat fetuses at 14 days' gestation were dispersed into a single-cell suspension by physical disruption and collagenase digestion. Pluripotent stem cells were characterized and partially purified by a combination of monoclonal antibodies. These included CD71 (anti-transferrin receptor, MRC-Oχ26, used for rosetting), Cdw90 (anti- Thy-1, MRC-Oχ7), and the newly described MRC-Oχ82 (reacting with myeloid cells in peritoneal exudate), employed in FACS sorting. Enrichment was monitored by long-term reconstitution of lethally irradiated congenic rats genetically distinguishable from the donor by an allelomorphic variant of the CD45 cell-surface antigen. At intervals from 3 months to 1 year, lymph-node cells and peritoneal exudate cells were biopsied for analysis by two-color flow cytometry-one color to determine donor origin, the other to identify Th cell (CD4+), Tc cell (CD8+), B cell (sIg+ or CD45RC+ ), neutrophil (Oχ82 + or Oχ43- ), and macrophage (Oχ43+) compartments. The degree of chimaerism was taken as the read out of stem-cell activity. No significant differentials between lymph-node and peritoneal exudate chimaerisms were detected in any of the recipients; therefore, the enrichment procedure revealed only pluripotent cells, not stem cells of restricted potency. All recovered stem-cell activity was in the Oχ26(CD71)-negative, Oχ7(CDw90)-positive, Oχ82-positive fraction. In the optimum case, an enrichment of very roughly 200-fold in cell-for-cell activity was obtained.

Rat bone-marrow colony-forming units in the spleen (CFUs-12) were found to lack the surface antigens recognized by the monoclonal antibodies CD53 (MRC-Oχ44), MRC-Oχ39, MRC-Oχ59, and 144.2.15. These would provide a strategy for their enrichment by depletion.