Simple Tissue-Culture Technique for Quantitating Free Migration of Reticulo-endothelial Cells

Abstract

This paper (1) describes a method for short-term, reproducible, quantitative measurement of the effects of experimental procedures on the migratory powers of reticuloendothelial cells in culture. Although splenic tissue was used in the preliminary work, the technique is adaptable for observations on mature and precursor motile cells in bone marrow and other organs.A 5-mm length of woven Fiberglas sleeving is prepared by repeated washing in xylol, alcohol, water, dilute HC1, NH4OH, and double-distilled water. Its lower 1.5 mm is made into a well-type receptacle by filling its interstices and the lower end with Diatex. (Diatex is a toluol solution of a transparent acrylic acid, plastic compound which hardens on drying and which can be autoclaved or sterilized in a heat oven. It has proved to be nontoxic in over 1200 tissue cultures. )The prepared sleeving is attached along its length to a piece of cover glass (Fig. 1). In the developmental stage of this project, half of a spleen from a 12day chick embryo, removed with the usual aseptic precautions, was used as test material. This piece of tissue is slipped into the open end of the Fiberglass sleeving, which with its attached cover glass is then placed in a Kahn tube containing 0.7 ml of supernatant, which consists of 0.25 percent human serum protein in Hanks' balanced salt solution. The Kahn tube is closed with a sterile serum-type rubber stopper containing a No. 20 hypodermic needle (fitted at its upper end with a cotton-plug bacterial filter).Any free, nonmigratory, dead or damaged cells from the fragment of tissue LO W o Z tq This paper (1) describes a method for short-term, reproducible, quantitative measurement of the effects of experimental procedures on the migratory powers of reticuloendothelial cells in culture. Although splenic tissue was used in the preliminary work, the technique is adaptable for observations on mature and precursor motile cells in bone marrow and other organs.A 5-mm length of woven Fiberglas sleeving is prepared by repeated washing in xylol, alcohol, water, dilute HC1, NH4OH, and double-distilled water. Its lower 1.5 mm is made into a well-type receptacle by filling its interstices and the lower end with Diatex. (Diatex is a toluol solution of a transparent acrylic acid, plastic compound which hardens on drying and which can be autoclaved or sterilized in a heat oven. It has proved to be nontoxic in over 1200 tissue cultures. )The prepared sleeving is attached along its length to a piece of cover glass (Fig. 1). In the developmental stage of this project, half of a spleen from a 12day chick embryo, removed with the usual aseptic precautions, was used as test material. This piece of tissue is slipped into the open end of the Fiberglass sleeving, which with its attached cover glass is then placed in a Kahn tube containing 0.7 ml of supernatant, which consists of 0.25 percent human serum protein in Hanks' balanced salt solution. The Kahn tube is closed with a sterile serum-type rubber stopper containing a No. 20 hypodermic needle (fitted ...

Workers and alate queens of Solenopsis geminata share qualitatively similar but quantitatively different venom alkaloid chemistry

“…Queens apply their venoms over eggs as they are laid, presumably for protecting eggs from entomopathogenic fungi (Vander Meer and Morel, 1995;Tschinkel, 2006). These alkaloids in queens may not necessarily function as defensive compounds as in workers (Blum et al, 1958;Javors et al, 1993), but conceivably as queen pheromones. Venom alkaloids on the surface of eggs may play a role in advertising the presence and fertility status of queens (Vander Meer and Morel, 1995;Eliyahu et al, 2011).…”

Characterization of the allergen Sol gem 2 from the fire ant venom, Solenopsis geminata

“…The toxicity of the venom is thought to be due to solenopsins and methyl-, alkylor alkenyl-substituted piperidines (7-9). The venom has cytotoxic, insecticidal, antibiotic and antimicrobial properties as well (7,(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19). The protein components of venom, which are wellknown allergens, are believed to play important roles in accelerating the spread of solenopsins to nearby cells and tissues.…”

Lethality of Red Imported Fire Ant Venom to Argentine Ants and Other Ant Species

“…Milking venom into a capillary tube placed on the sting was first described by Blum et al (1958), and later widely used in obtaining pure venom for compositional analyses or bioassays (MacConnell et al 1970;1971;Brand et al 1972Brand et al , 1973bMacConnell et al 1976;Blum 1992;Greenberg et al 2008;Lai et al 2008Lai et al , 2009Lai et al , 2012. In this way, milligram quantities of pure venom can be obtained for direct use.…”