Balanced salt solutions

Historically, the development of animal cell culture systems has been dependent upon the development of new types of tissue culture media. Mouse L cells and HeLa cells were developed using a balanced salt solution supplemented with blood plasma, an embryonic tissue extract, and/or serum. In 1955 Eagle developed a nutritionally defined medium, containing all of the essential amino acids, vitamins, cofactors, carbohydrates, salts, and small amounts of dialyzed serum (Table 1). He demonstrated that this minimal essential medium (MEM) supported the long-term growth of mouse L and HeLa ceils. Eagle s MEM was so well defined that the omission of a single essential nutrient eventually resulted in the death of these animal cells in culture. 

HBBS Hank s balanced salt solution HCA Hypertonic citrate H-CAM Hyaluronic acid cell adhesion molecule HDC Histidine decarboxylase... 

HBSS Hanks balanced salt solution (GibcoBRL, Eggenstein, Germany). 

Starvation and incubation with labeled dextran For this method, cells are incubated under starvation [Earle s balanced salt solution (BBSS) medium] conditions for two hours. To label early endocytic compartments, cells are subsequently incubated with 0.5 mg/mL dextran tetramethylrhoda-mine for five minutes. To label late endosomes, cells are washed and further incubated for an additional 10 minutes (130). 

Sterile Petridish (2) Sterile balanced salt solution (3) 0.25% Trypsin in PBS (4) Growth media V J... 

Transfer the tissue to a sterile balanced salt solution on ice. 

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

Add Hank s balanced salt solution (room temperature) to the mixture to bring the vol to 50 mL, and centrifuge the cells for 10 min at 500g. 

Decant the supernatant, and wash the cells with Hank s balanced salt solution two more times. The cells should be relatively free of contaminating red blood cells and platelets after these three washing steps. 

At various time intervals, stop endocytosis with cold (4°C) PBS or balanced salt solution. 

The cores are subsequently placed in the sheer, and the slicing procedure is performed by advancing the core over an oscillating knife in a controlled environment (Figure 12.1). Cold (4°C) Krebs-Henseleit buffer (pH = 7.4, saturated with 95% O2 and 5% CO2) supplemented with 25 mM glucose is commonly used in preparing the slices [35,38 0], but Williams medium E [41], Earle s balanced salt solutions [37], Sacks preservation medium [42] and V-7 preservation buffer [43,44] are also used. 

Concomitantly, the removed volume has to be replaced to avoid ocular hypotension. This volume replacement is performed with a balanced salt solution. At the end of the vitrectomy, the vitreous gel has been removed—ideally completely—and the vitreous cavity is filled now with balanced salt solution. 

It must be mentioned here that some of the results discussed in Sect. 4.3 were obtained with cell suspensions in Hanks -balanced salt solution (HBSS) in the absence of plasma proteins. The present author believes that the random-network concept can be applied to the events which happen to lymphocytes, platelets or erythrocytes when they come into direct contact with hydrated material surfaces in the absence of interventing protein. 

Grow the fibroblasts in Ham F-10 medium with 10% fetal calf serum (FCS). Harvest the cells, seed at a density of 30,000 cells and grow to near confluency on cover slips in 6-well plates in Ham F-10 medium with 10% FCS. Remove the medium and wash the cells three times with 2 ml Hanks balanced salt solution. Fix the cells with 3 ml fixative. Seal the plate with tape and store in the refrigerator at 4 C until staining. 

Hank s Balanced Salt Solution (HBSS), pH 7.4 (Gibco, Paisley, Scotland). Store at 15-30°C... 

The suspensions are centrifuged and washed twice in 3% FBS/PBS. They are again centrifuged and washed twice in 3% FBS in PBS with 0.1% Triton X-100. The pellets are resuspended and incubated with 0.5 ml of 1% FBS containing propidium iodide (10 pg/ml Calbiochem, San Diego, CA) and RNAse (1 mg/ml) in Hank s balanced salt solution (without phenol red) for 30 min at 37°C. The samples are analyzed on a Coulter Electronics XL-MCL or a Becton Dickinson FACScan flow cytometer with standard argon ion laser excitation and filter configuration for the FTTC/propidium iodide dye combination. 

Hank s balanced salt solution (HBSS JRH Biosciences). 

Tiypsin-ethylenediamine tetraacetic acid (EDTA) in Hank s balanced salt solution, without calcium or magnesium (Invitrogen, Carlsbad, CA). 

Metastasis 2. Medium for tumor cell culture, e.g., Minimal Essential Medium Eagle (MEM) (Gibco, Invitrogen) 3. 10X Trypsin/EDTA dilute to final concentration lx with PBS (Sigma-Aldrich) 4. Standardized tumor cell suspension, radiolabeled 5. Hank s balanced salt solution, Ca++ and Mg++ free (CMF-HBSS) (Sigma-Aldrich, H-2387) 6. Trypan blue stain 7. Mouse vice 8. Heat lamps... 

Metastasis 2. Hank s balanced salt solution, Ca++ and Mg++ free (CMF-HBSS) 3. Methoxyfluorane or xylazine/ketamine 4. Nail and hair removal cream... 

To avoid clumping of cells, the cell preparations must be free of serum and the tumor cells should be injected in a Ca++- and Mg++-free balanced salt solution (BSS), which also serves to decrease clumping. 

Add calcium and magnesium free-balanced salt solution (CMF-HBSS) ( 5 ml/25 cm2) to the side of the flask opposite the cells so as to avoid dislodging cells, rinse cells, and discard rinse (see Note 5). 

Bone marrow is placed into a balanced salt solution containing preservative-free heparin and a single cell suspension prepared by passing the marrow through a fine wire mesh. Alternatively, a small Potter-Elvehjem tissue homogenizer (Hll) may be used. The nucleated cell count is adjusted to 5-10 X 106 ml of balanced salt solution and deoxyuridine added. Following incubation, 5 p,Ci of tritiated thymidine is added and the mixture incubated for an additional hour. The cells are then washed and the nucleated cell count determined. A 0.1 ml aliquot is then placed on a filter paper disk and dried. The activity of the dried disks is then measured in a scintillation... 

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