Preparation and characterization of viable epithelial cells from rabbit distal colon

Journal Article · Fri Apr 01 00:00:00 EST 1988 · American Journal of Physiology; (USA)

OSTI ID:7016121

Kaunitz, J D ^[1]

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1. West Los Angeles Veterans Administration Hospital, CA (USA) Univ. of California, Los Angeles (USA)

A preparation of viable epithelial cells, suitable for transport studies, was prepared from rabbit distal colon. Enzymatic digestion of scraped mucosa liberated a population of intact colonic glands that were dissociated into cells by gentle agitation in culture medium. Metabolism of ({sup 14}C)glucose was constant over 2 h and was inhibited 70% by 1 mM ouabain. Cells were loaded with the trapped, pH-sensitive fluorescent dye 2{prime}-7{prime}-bis(carboxyethyl)-5(6)-carboxyfluorescein acetoxy methyl ester and leaked 0.1% of the dye per minute. Cell pH was 7.21 {+-} 0.03 in pH 7.4. Ringer medium. Intracellular potassium concentration, as measured by a nigericin null-point technique, was 128 {+-} 8 mM. Plasma membrane potential measured with the potential-sensitive fluorescent dye 3,3-dipropylthiadicarboxycanine iodide was -52 {+-} 2 mV. Recovery of intracellular pH in acid-loaded cells occurred after exposure to sodium-containing medium and was inhibited by 5 {times} 10{sup {minus}4} M amiloride. It is concluded that viable epithelial cells can be prepared from rabbit distal colon with relative simplicity and in high yield. These cells are suitable for measurement of intracellular pH and membrane potential and are thus a convenient model system for study of colonic cell physiology.

• OSTI ID: 7016121
• Journal Information: American Journal of Physiology; (USA), Vol. 254:4; ISSN 0002-9513
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
CARBON 14 COMPOUNDS
METABOLISM
CELL MEMBRANES
ELECTRIC POTENTIAL
EPITHELIUM
CYTOLOGY
GLUCOSE
CATIONS
DIURETICS
FLUORESCEIN
LARGE INTESTINE
PH VALUE
POTASSIUM COMPOUNDS
RABBITS
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ANIMALS
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BODY
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CARBOXYLIC ACIDS
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HEXOSES
HYDROXY ACIDS
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IONS
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MAMMALS
MEMBRANES
MONOSACCHARIDES
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550301* - Cytology- Tracer Techniques