Schot R, van Asselt E, van Mastrigt R
Department of Urology, Erasmus University Rotterdam, The Netherlands.
Urol Res. 1993 Jan;21(1):49-53. doi: 10.1007/BF00295192.
The present study describes a method for isolating single smooth muscle cells from pig urinary bladder using a continuous resuspension device. Low concentrations of collagenase and papain were sufficient to obtain a high yield of viable smooth muscle cells, which remained viable for about 3-4 h as tested with fluorescein diacetate. Addition of fetal calf serum increased the lifespan of the isolated cells and the percentage of contractile smooth muscle cells, but caused spontaneous shortening of the cells. The length and volume of the isolated smooth muscle cells depended on the calcium concentration used in the isolation buffer solution. The isolated muscle cells were apparently relaxed if a calcium concentration less than 1.0 mmol/l was used in the isolation medium. In higher calcium concentrations the isolated cells were significantly shorter, probably as a result of a contraction caused by mechanical stimulation of the cells during the isolation procedure.
本研究描述了一种使用连续重悬装置从猪膀胱中分离单个平滑肌细胞的方法。低浓度的胶原酶和木瓜蛋白酶足以获得高产率的存活平滑肌细胞，用荧光素二乙酸酯测试表明这些细胞可存活约3 - 4小时。添加胎牛血清可延长分离细胞的寿命以及收缩性平滑肌细胞的百分比，但会导致细胞自发缩短。分离的平滑肌细胞的长度和体积取决于分离缓冲溶液中使用的钙浓度。如果在分离培养基中使用低于1.0 mmol/l的钙浓度，分离的肌肉细胞明显松弛。在较高钙浓度下，分离的细胞明显更短，这可能是由于在分离过程中细胞受到机械刺激而引起收缩的结果。
