Preparation and characterization of viable epithelial cells from rabbit distal colon

Journal Article · Fri Apr 01 04:00:00 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), Journal Name: American Journal of Physiology; (USA) Vol. 254:4; ISSN 0002-9513; ISSN AJPHA
• Country of Publication: United States
• Language: English

Related Subjects

550301* -- Cytology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
ALKALI METAL COMPOUNDS
ANIMAL TISSUES
ANIMALS
AROMATICS
BIOLOGY
BODY
CARBOHYDRATES
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
CATIONS
CELL CONSTITUENTS
CELL MEMBRANES
CHARGED PARTICLES
CYTOLOGY
DIGESTIVE SYSTEM
DIURETICS
DRUGS
DYES
ELECTRIC POTENTIAL
EPITHELIUM
FLUORESCEIN
GASTROINTESTINAL TRACT
GLUCOSE
HEXOSES
HYDROXY ACIDS
HYDROXY COMPOUNDS
INTESTINES
IONS
LABELLED COMPOUNDS
LARGE INTESTINE
MAMMALS
MEMBRANES
METABOLISM
MONOSACCHARIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PH VALUE
PHENOLS
POLYPHENOLS
POTASSIUM COMPOUNDS
RABBITS
SACCHARIDES
TISSUES
VERTEBRATES