Isolation of viable mouse primordial germ cells by antibody-directed flow sorting

The germ line represents a cell type of unique interest in mammals because it retains complete genotypic totipotency while undergoing significant phenotypic differentiation. Analysis of the mechanism that underlies the maintenance of this totipotency requires the ability to isolate and study all stages of the germ cell lineage. The primordial germ cells (PGC) are the earliest identifiable germ cells in the embryo. It has not previously been possible to isolate PGC in sufficient numbers and purity to facilitate biochemical and/or molecular analysis. We report here that the use of a monoclonal antibody in combination with flow cytometry does permit the isolation of reasonably large and pure yields of viable mouse PGC.     

Thermal behavior of HeLa and KB cells in suspension and attached to glass

HeLa S-3 and KB cells were grown in a LKB Batch Microcalorimeter under a variety of nutrient medium conditions and mixing intervals. These conditions produced rather large apparent endothermic and exothermic responses on mixing that could be correlated with the presence of suspended cells (unattached) as well as cells attached to the glass calorimeter vessel. Cells capable of being resuspended upon mixing of the calorimeter vessel produces first an endothermic followed by an exothermic signal while attached cells produced only an apparent endothermic response. The exothermic response is believed to be associated with increased metabolic heat on suspending the cells followed by partial suppression of the steady state metabolic heat on cell settling. Rates of cell settling correlated well with the rate of decay of the exothermic signal. The rapid appearance of endothermicity on mixing suggests it is associated with rapid events such as binding of nutrients to cell surfaces. The response in the endothermic direction on mixing is discussed in terms of the disruption of mechanisms which tend to exclude nutrients from the surface of the cell.     

A function for the lck proto-oncogene

Proto-oncogenes encode products that comprise a select group of cellular regulatory proteins whose mutation or aberrant expression can result in oncogenic transformation. With the exception of certain growth factors and their receptors, the definition of normal functions for most proto-oncogene products has been elusive. The discovery that a member of the src-family of tyrosine protein kinases (p56lck) is associated with both the CD4 and CD8 T-lymphocyte surface glycoproteins provides the first clue to understanding the potential physiological functions of this family of proto-oncogenes.