Mycoplasma hominis: growth, reproduction, and isolation of small viable cells

Improved methods for studying the growth of Mycoplasma hominis (ATCC 14027) have been developed, involving modified growth conditions and preparation of the organisms under minimally distorting conditions. Cells so prepared from batch cultures show relatively uniform exponential growth and appear to be dividing by binary fission; but pleomorphic forms appear upon further incubation. Similar behavior was demonstrated by another laboratory-adapted strain and by three clinical isolates, and therefore seems characteristic of the species. The pleomorphic populations contain small forms having diameters within the 100- to 250-nm size range reported for "elementary bodies." Such forms were isolated from this strain of M. hominis by sequential filtration using gravity alone, after cell aggregates were dispersed by Pronase treatment. Of the small bodies which traversed membranes of 220-nm pore size, a negligible number grew in liquid or on solid media, suggesting that these were not essential reproductive units in a life cycle, but involution forms due to growth in an altered environment.