Pearce F L, Behrendt H, Blum U, Poblete-Freundt G, Pult P, Stang-Voss C, Schmutzler W
Agents Actions. 1977 Mar;7(1):45-56. doi: 10.1007/BF01964880.
A method is described for the dissociation of guinea pig lung and mesentery into their component cells. The method comprises incubation of the tissues with the enzyme pronase in Ca++- and Mg++-free saline followed by mechanical dispersion and yields on average 3-8% free mast cells. These cells are morphologically intact and viable. They retain an active sensitization or can be passively sensitized. The levels of cAMP and cGMP in the whole cell suspension respond to catecholamine, histamine or cholinergic stimulation in a way similar to that observed in the corresponding undispersed tissue. Although the separation of guinea pig mast cells from other cells proved to be more difficult than the purification of rat mast cells, an inrichment of greater than 50% has been achieved. The mast cells isolated by this method proliferate in vitro and have been cultured successfully for several months.
本文描述了一种将豚鼠肺和肠系膜解离成其组成细胞的方法。该方法包括将组织与链霉蛋白酶在无钙和无镁的盐溶液中孵育,随后进行机械分散,平均可得到3 - 8%的游离肥大细胞。这些细胞形态完整且具有活力。它们保留了活跃的致敏作用,或者可以被被动致敏。全细胞悬液中cAMP和cGMP的水平对儿茶酚胺、组胺或胆碱能刺激的反应方式与在相应未分散组织中观察到的相似。尽管事实证明从其他细胞中分离豚鼠肥大细胞比纯化大鼠肥大细胞更困难,但已实现了超过50%的富集。通过这种方法分离的肥大细胞在体外增殖,并已成功培养了数月。