Metabolic and structural viability of precision-cut rat lung slices in culture

Abstract

1. The principal objective was to evaluate the functional and structural integrity of precision-cut rat lung slices in culture over 72 h.2. Lung slices metabolized 7-ethoxycoumarin in a time-dependent fashion, the major metabolites being the sulphate and glucuronide of 7-hydroxycoumarin with very low levels of the free compound. Prior treatment of rats with b-naphthoflavone elevated markedly the rate of metabolism. The optimum slice thickness, as exemplified by the metabolism of 7-ethoxycoumarin, was about 600 mm.3. Lung slices retained metabolic viability towards 7-ethoxycoumarin for 8 h, but after this point a marked decline in metabolic activity was noted. However, very low levels of activity were still evident following a 72 h incubation.4. Morphological examination of lung slices revealed nuclear degeneration and loss of tissue architecture following 24 h incubation. When cellular integrity was assessed using lactate dehydrogenase, a time-dependent leakage was evident with maximum loss occurring within 24 h; longer incubations did not result in further leakage. 5. It is concluded that precision-cut rat lung slices, of 600 mm thickness, can be maintained metabolically viable in culture for some 8 h.

Modulation of rat pulmonary carcinogen-metabolising enzyme systems by the isothiocyanates erucin and sulforaphane

“…Precision-cut rat lung slices were prepared as previously described [15]. Briefly, lungs were perfused intratracheally with agarose (0.75% v/w) at 37 °C.…”

“…The studies reported here were conducted in vitro utilising precisioncut lung slices [15] The principal advantages of this in vitro system are that it maintains tissue architecture and cell-cell communication, and allows the facile use of a wide range of concentrations of chemicals, that would otherwise necessitate large animal numbers for commensurate in vivo studies [16]. It is particularly suited to the lung, a heterogeneous tissue composed of many different cell types that differ markedly in their carcinogen-metabolising enzyme profile [17].…”

Up-regulation of CYP1A/B in rat lung and liver, and human liver precision-cut slices by a series of polycyclic aromatic hydrocarbons; association with the Ah locus and importance of molecular size

“…were prepared from cylindrical cores (8 mm) as described for the liver (Umachandran et al, 2004(Umachandran et al, , 2006. A pre-incubation of 60 minutes was carried out.…”

“…The advantages of adopting this in vitro system, in comparison with other systems such as subcellular fractions and primary hepatocytes, have frequently been pointed out in reviews (de Kanter et al, 1999;Lerche-Langrand and Toutain, 2000). Slices have been prepared from a number of tissues including liver (Hashemi et al, 1999), lung (Umachandran et al, 2004), intestine and colon (van de Kerkhof et al, 2005).…”

Differential response of human and rat epoxide hydrolase to polycyclic aromatic hydrocarbon exposure: Studies using precision-cut tissue slices

“…For the production of lung slices, animals were killed by an overdose of sodium pentobarbital, and lungs were perfused intratracheally with agarose (0.75% v/w) at 37 °C. Agar was allowed to solidify, and lung slices (600 μm) were prepared from cylindrical cores (8 mm) as described for the liver [17]. For incubations exceeding 24 h, slices were placed in fresh medium every 24 h. Three different slice pools, comprising 4-10 slices, were used per time point.…”