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目的建立一种简便有效的离体肺组织切片孵育方法。方法利用肺组织精细切片技术制备肺组织切片 ,以 0 .5mLKrebs Henseleit(K H)缓冲液作为孵育液 ,在 37℃培养箱内分别孵育 1、2、3、4h ,测定肺片的四甲基偶氮唑盐 (MTT)比色、乳酸脱氢酶 (LDH)释放率和三磷酸腺苷 (ATP)含量 ,以反映肺组织活性。结果肺片在 37℃培养箱内孵育 1、2、3、4h后MTT比色、LDH释放率和ATP含量均无统计学差异 (P >0 .0 5 )。结论应用 0 .5mLKrebs Henseleit(K H)缓冲液作为孵育液、在 37℃培养箱对肺片进行孵育时组织活性没有明显变化 ,可作为一种简便有效的孵育方法用于离体肺片研究
Objective To establish a simple and effective incubation method of isolated lung tissue sections. Methods Pulmonary tissue sections were prepared by fine slicing technique of lung tissue. After incubation with 0.5 mL Krebs Henseleit (KH) buffer for 1, 2, 3 and 4 h in a 37 ° C incubator, the tetramethyl couple MTT colorimetric, lactate dehydrogenase (LDH) release rate and adenosine triphosphate (ATP) content were measured to reflect lung tissue activity. Results MTT colorimetric assay, LDH release rate and ATP content showed no statistical difference (P> 0.05) after incubation for 1, 2, 3, 4 h in 37 ℃ incubator. Conclusions Using 0.5 mL Krebs Henseleit (K H) buffer as incubation solution, there is no significant change in the tissue activity when incubated in the 37 ° C incubator, which can be used as a convenient and effective method for the isolation of lung slices