Permeabilization of cells of hemopoietic origin by extracellular ATP4-: elimination of osteoclasts, macrophages, and their precursors from isolated bone cell populations and fetal bone rudiments.
Skeletal tissues contain, apart from cells of the osteogenic and chondrogenic lineage, cells of hemopoietic origin, e.g., macrophages, osteoclasts, and their precursors. In the present study we examined the sensitivity for extracellular ATP4- of the above-mentioned cell types in freshly isolated, bone-derived cell populations and in explanted fetal metatarsal bones. Cells of hemopoietic origin reacted to the presence of ATP4- with an increased permeability for impermeant cytotoxic molecules, e.g., ethidium bromide (EB), thiocyanate (KSCN), and an increased non-ion selective membrane conductance. As a consequence, these cells could be killed by a short treatment with adenosine-5' triphosphate (ATP)+KSCN. On the other hand, cells of nonhemopoietic origin (e.g., osteoblasts, chondrocytes) were found to be insensitive to ATP4- in this respect. These cells survived the treatment without apparent damage to their alkaline phosphatase activities, osteogenic potentials, and osteoclast induction capacities. The elimination of the endogenous cells of hemopoietic origin from bone tissue or cell populations derived therefrom offers the possibility to study the properties and functions of osteogenic or chondrogenic cells without interference by the presence of cells of hemopoietic origin. It also allows the study of interactions between osteogenic cells and selected cell populations of hemopoietic origin in coculture experiments.
Department of Cell Biology and Histology, University of Leiden, The Netherlands.
W E
WE
Modderman
A F
AF
Weidema
T
T
Vrijheid-Lammers
A M
AM
Wassenaar
P J
PJ
Nijweide
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001842:Bone and Bones / Q000166:cytology*; D002463:Cell Membrane Permeability / Q000187:drug effects; D002469:Cell Separation; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects; D006413:Hematopoietic System / Q000166:cytology*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D009499:Neutral Red; D010010:Osteoclasts / Q000166:cytology / Q000187:drug effects
D009499:Neutral Red; D000255:Adenosine Triphosphate