Extraction of leucocytes from human decidua. A comparison of dispersal techniques.
Functional studies of human utero-placental tissues have been limited by poor characterisation of the morphology and antigenic phenotype of the cells under investigation. The present study documents the effect of dispersal methods on the viability and cellular composition of cell suspensions prepared from decidualised endometrium in early human pregnancy. First trimester decidua was subjected to both mechanical disaggregation and digestion with various enzyme combinations in an attempt to optimise the yield of infiltrating decidual leucocytes. Cell types were characterised with monoclonal antibodies using an alkaline phosphatase immunolabelling method. Mechanical disaggregation resulted in suspensions containing many large decidual cells and much cell debris but few leucocytic cells. Overall viability was low, although the viability of small leucocytes common antigen-bearing cells remained high. Enzymic digestion yielded cell suspensions rich in leucocytes with high viability and minimal contamination by other cell types. Collagenase produced a high yield of leucocytes with high viability and minimal disruption of surface antigens in contrast with pronase which caused extensive antigenic loss. The disaggregation method determines the yield of bone marrow derived cells in decidual cell suspensions and the extent of contamination by true decidual cells and epithelial cells.
Ritson
A
A
Department of Pathology, University of Leeds, U.K.
Bulmer
J N
JN
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis; D002469:Cell Separation / Q000379:methods; D003656:Decidua / Q000166:cytology*; D004798:Enzymes; D004848:Epithelium / Q000276:immunology; D005260:Female; D006801:Humans; D007962:Leukocytes / Q000166:cytology*; D008264:Macrophages / Q000276:immunology; D011247:Pregnancy
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D000954:Antigens, Surface; D004798:Enzymes