title: Immunomagnetic isolation of fetal rat gonocytes.
abstract: OBJECTIVE
  An efficient method to obtain highly enriched populations of viable gonocytes from rat embryos at Day 18 and Day 20 postcoitum (pc) is described.
  
  
  METHODS
  Single-cell suspensions with high cell yield were obtained by a collagenase/ trypsin digestion of the decapsulated testis. The gonocytes were purified by a direct immunoseparation technique, using magnetizable beads coated with rat anti-mouse immunoglobulin M (IgM) and a monoclonal antibody 4B6.3E10, which specifically reacted with a differentiation antigen on the fetal germ cells.
  
  
  RESULTS
  Populations of 8.3 +/- 2.7 (x10(3); 18 days pc) or 1.2 +/- 0.25 (x10(4); 20 days pc) viable gonocytes per testis with purities of 91 +/- 6.5% and 92 +/- 4.3%, respectively, as determined by Nomarski microscopy were obtained.
  
  
  CONCLUSIONS
  The cells were successfully used for culture studies and as starting material for the investigation of gene expression.
authors.affiliation: Department of Functional Morphology, Veterinary School, Utrecht University, The Netherlands.
authors.forename: R
authors.initials: R
authors.lastname: van den Ham
authors.forename: A M
authors.initials: AM
authors.lastname: van Pelt
authors.forename: M P
authors.initials: MP
authors.lastname: de Miguel
authors.forename: P J
authors.initials: PJ
authors.lastname: van Kooten
authors.forename: N
authors.initials: N
authors.lastname: Walther
authors.forename: F M
authors.initials: FM
authors.lastname: van Dissel-Emiliani
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D005260:Female; D007075:Immunoglobulin M; D018189:Immunomagnetic Separation / Q000379:methods*; D008297:Male; D011247:Pregnancy; D012333:RNA, Messenger / Q000302:isolation & purification; D051381:Rats; D017208:Rats, Wistar; D013737:Testis / Q000737:chemistry / Q000166:cytology*
chemical_list: D000911:Antibodies, Monoclonal; D007075:Immunoglobulin M; D012333:RNA, Messenger