Shleeva M O, Mukamolova G V, Telkov M V, Berezinskaia T L, Syroeshkin A V, Biketov S F, Kaprel'iants A S
Bach Institute of Biochemistry, Russian Academy of Sciences, Leninskii pr. 33, Moscow, 117071 Russia.
Mikrobiologiia. 2003 Jan-Feb;72(1):76-83.
Nonculturable cells were found to occur in populations of Mycobacterium tuberculosis cells during the long post-stationary phase. These cells were small (0.6-0.8 micron) ovoid and coccoid forms with intact cell walls and negligible respiratory activity, which allows them to be regarded as dormant cells. Nonculturable cells were characterized by low viability after plating onto solid medium; a minor part of the population of these cells could be cultivated in liquid medium. Cell-free culture liquid of an exponential-phase Mycobacterium tuberculosis culture or the bacterial growth factor Rpf exerted a resuscitating effect, increasing substantially the growth capacity of the nonculturable cells in liquid medium. During resuscitation of nonculturable cells, a transition from ovoid to rodlike cell shape occurred. At early stages of resuscitation, ovoid cells formed small aggregates. The recovery of culturability was associated with the formation of rod-shaped cells in the culture. The data obtained demonstrate the in vitro formation of dormant cells of Mycobacterium tuberculosis, which do not grow on solid media but can be resuscitated in liquid medium under the effect of substance(s) secreted by actively growing cells.
在结核分枝杆菌细胞群体的长期稳定后期发现了不可培养细胞。这些细胞呈小的（0.6 - 0.8微米）卵形和球形，细胞壁完整，呼吸活性可忽略不计，这使得它们可被视为休眠细胞。不可培养细胞的特征是接种到固体培养基后活力较低；这些细胞群体中的一小部分可以在液体培养基中培养。指数生长期结核分枝杆菌培养物的无细胞培养液或细菌生长因子Rpf具有复苏作用，可显著提高不可培养细胞在液体培养基中的生长能力。在不可培养细胞复苏过程中，细胞形状从卵形转变为杆状。在复苏早期，卵形细胞形成小聚集体。可培养性的恢复与培养物中杆状细胞的形成有关。所获得的数据证明了结核分枝杆菌休眠细胞的体外形成，这些细胞在固体培养基上不生长，但在活跃生长细胞分泌的物质作用下可在液体培养基中复苏。
