A method for isolation and partial purification of Trichinella spiralis nurse cells.
Invasion of vertebrate muscle cells by larvae of Trichinella spiralis is accompanied by redifferentiation of the host myofiber into a novel structure called the nurse cell. The nurse cell protects and nurtures the enclosed parasite during its long stay in host muscle. It is anatomically independent of the surrounding uninfected muscle cells and can be isolated from host tissue by mechanical or enzymatic means. Current methods employed for this purpose have yielded only small numbers of nurse cells. An apparatus designed to isolate large numbers of nurse cells and a method for removal of all free larvae and most host muscle debris is described. Homogenization and trypsin digestion of muscle tissue was followed by passage of muscle/parasite suspensions maintained at 37 C through a jacketed glass column fitted with a 40-mesh stainless steel screen at the top and a Nitex screen with 150-microns-diameter pores at the bottom. Nurse cells were retained by the Nitex screen. Density gradient centrifugation using Percoll removed all free larvae and most contaminating muscle debris from nurse cell suspensions. The large quantities of nurse cells made available by this method will allow evaluation of the molecular biology, nutrition, biochemistry, and metabolism of the enclosed parasite and of the Trichinella-modified host muscle cell.
Montgomery
J
J
Center for Parasitology, University of Texas at Arlington 76019, USA.
Feldman
A
A
Despommier
D D
DD
Stewart
G L
GL
Haehling
E
E
D000818:Animals; D002469:Cell Separation / Q000379:methods; D002499:Centrifugation, Density Gradient; D007814:Larva / Q000502:physiology; D051379:Mice; D008813:Mice, Inbred ICR; D018482:Muscle, Skeletal / Q000166:cytology / Q000469:parasitology*; D017160:Trichinella spiralis / Q000302:isolation & purification / Q000502:physiology*; D014357:Trypsin / Q000378:metabolism
D014357:Trypsin