Edmondson Diane G, Norris Steven J, De Lay Bridget D
University of Texas Health Science Center at Houston - McGovern Medical School;
University of Texas Health Science Center at Houston - McGovern Medical School.
J Vis Exp. 2025 Jan 24(215). doi: 10.3791/66880.
For over a century, Treponema pallidum subsp. pallidum, the spiral-shaped bacterium that causes syphilis, could only be propagated by inoculation and harvest of the organisms from rabbit testes. In 2018, we described a method to continuously cultivate T. pallidumin vitro. This system utilizes co-culture with rabbit epithelial cells (Sf1Ep cells) in a serum-containing tissue culture medium called TpCM-2. The T. pallidum doubling time in culture is similar to that estimated to occur during natural infection (about 33-45 h). The organism can be cultured continuously with a standard passage time of 1 week in a low oxygen (1.5%) environment at 34 °C. This article contains the protocols for culturing T. pallidum, methods for growing and maintaining the required tissue culture cells, and the technique for generating isogenic strains by limiting dilution. The ability to grow T. pallidum in vitro provides new experimental avenues to study and understand this enigmatic organism.
一个多世纪以来，引起梅毒的螺旋形细菌梅毒螺旋体苍白亚种只能通过从兔睾丸接种和收获该生物体来繁殖。2018年，我们描述了一种在体外连续培养梅毒螺旋体的方法。该系统利用在一种名为TpCM - 2的含血清组织培养基中与兔上皮细胞（Sf1Ep细胞）共培养。梅毒螺旋体在培养中的倍增时间与自然感染期间估计的倍增时间相似（约33 - 45小时）。该生物体可以在34°C的低氧（1.5%）环境中以1周的标准传代时间连续培养。本文包含梅毒螺旋体培养方案、培养和维持所需组织培养细胞的方法以及通过有限稀释产生同基因菌株的技术。在体外培养梅毒螺旋体的能力为研究和了解这种神秘生物体提供了新的实验途径。
