Hank’s-buffered salt solution

Cell washing solution This should be an isotonie, protein-free solution at a neutral pH, sueh as phosphate-buffered saline or Hank s buffered salt solution. 

Hank s-buffered salt solution (HBSS) (Cambrex BioSciences, Inc., Walkersville, MD). 

Hank s balanced salt solution (Gibco-BRL, Grand Island, NY). Phosphate-buffered saline ([PBS] Gibco-BRL). 

The cerebral cortices of six rats were dissected and immediately placed in ice-cold Hank s balanced salt solution (HBSS) at pH7.5, supplemented with 50 qL/mL penicillin, 50 qg/mL streptomycin and 10 000 KIU/L aprotinin. The cortices were then rinsed twice with ice-cold HBSS and homogenized in 10 vol. of HBSS. The suspension was centrifuged at 2600 rev./min for 10 min at 4°C in a Hitachi CF7D2 centrifuge, and the pellet was homogenized in a buffer... 

To the cells previously washed with internalization media (step (h) of the previous protocol), 1 mL of sodium acetate 20niM in Hank s balanced salt solution (HBSS) is added, pH5.0 (see Table I.l for the composition of HBSS). It is also possible to use O.IM glycine buffer at pH2.8. 

Caco-2 monolayers are incubated at 37°C for 30 min in a C02 incubator with transport media (TM), prior to initiating a transport experiment. The TM for the apical side is made of Hank s balance salt solution (HBSS) containing lOmM HEPES buffer (pH 7.4) and 25 mM D-glucose (a recent study suggested that simulated intestinal fluid might be a better media to use in the apical compartment especially for highly lipophilic compounds [66,67], and this new media will be investigated in the future). 

Fig 1. Effect of incubation meditun and excess hormone on L-T and L-T/, uptake by mouse neuroblastoma cells (NBATA3) After 45 min preincubation confluent cells were incubated at 37°C for 2 h with 10 pM labeled thyroid hormone in the absence or presence of lOpM unlabeled hormone. Wells measuring 28.2 cm contained 4 ml of culture medium (RPMI 1640) or Hank s balanced salt solution. After washing with phosphate buffered saline +0.1% serum albumin, the cells were harvested and their radioactivity measured. Mean SEM of 3 experiments performed in duplicate. Experiments were performed under diminished light to avoid spontaneous deiodination especially of T, in RPMI due to riboflavin and folic acid. 

Figure 11.26 A schematic diagram of an apparatus used to obtain estimates of the passive permeability of a drug candidate across the intestinal mucosa using Caco-2 cells. A monolayer of Caco-2 cells is grown on a porous polyethylene terephthalate (PET) membrane (a so-called confluent monolayer of cells that grows only in two dimensions on such a substrate from an initial small inoculation). In the experiment the cells are submerged in Hanks s Balanced Salt Solution (HBSS) buffer (contains Na+, K+, CP, phosphate, glucose, and in some formulations also Ca +, Mg + and S04 ) the Caco-2 cell layer provides the only connection between an apical (donor) reservoir, into which the drug candidate is dosed, and a basolateral (receiver) reservoir. For the assay, aliquots are removed for analysis from the apical reservoir at Omin, and from both reservoirs at 120 min. Reproduced from Van Pelt, Rapid Commun. Mass Spectrom. 17, 1573 (2003), with permission of John Wiley Sons Ltd.

Rochester, N.Y.) as supporting electrolyte. Aqueous samples were run in phosphate buffered saline pH 7.5 (PBS), Hank s balanced salt solution (HESS) pH 7.5 or unbuffered water adjusted to pH 7.5. The electrolyte was 0.05 M KCl. 

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