Fractionation and characterization of the blast cell population of the mouse thymus by physical cell separation methods and ultrastructural analysis

• Published in: Leukemia research Vol. 3; no. 3; pp. 135 - 146
• Main Authors: Boniver, Jacques, Declève, Alain, Lepoint, Alain, Courtoy, Robert, Simar, Léon J.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 1979
• Subjects: Animals; cell separation; Cell Separation - methods; Centrifugation, Density Gradient; Chemical Fractionation; Chemical Precipitation; Female; leukemogenesis; Lymphocytes; lymphopoiesis; Mice; Mice, Inbred C57BL; Microscopy, Electron; mouse thymic blast cells; Thymus Gland - cytology; Thymus Gland - ultrastructure; ultrastructure; mouse thymic blast cells; lymphopoiesis; leukemogenesis; RadLV, Radiation leukemia virus; PBS, Phosphate buffered saline; Lb cell, Lymphoblast; BSS, Balanced salt solution; cell separation; RSN cell, Ring-shaped nucleolus cell; ultrastructure; BSA, Bovine serum albumin
• ISSN: 0145-2126; 1873-5835
• DOI: 10.1016/0145-2126(79)90011-0

Sedimentation at unit gravity and discontinous density gradients were used either separately or sequentially in an attempt to isolate cell preparations that are significantly enriched in one of the several subtypes of thymocytes present in the C57BL mouse thymus. Electron microscopy analysis of the cellular content of fractions obtained by such methods demonstrates that fractionation by 1g sedimentation allows the isolation of either pure preparations of small cortical lymphocytes or of fractions that are significantly enriched in blast cells, whereas density gradient centrifugation yields to the selection of the low to medium density lymphoblasts. A sequential separation procedure using both sedimentation at unit gravity and density gradient centrifugation was found to be the method of choice to isolate pure preparations of medullary lymphoid cells or to obtain fractions devoid of small lymphocytes and highly enriched in blast cells of the X subtype. These methods may thus serve as a basis for the selective isolation of putative target cells which are believed to be present in the thymus and to play a role in the pathogenesis of thymic lymphomas in C57BL mice.