A simple perfusion technique for isolation of maternal intervillous blood mononuclear cells from human placentae.
A noninvasive perfusion method for the recovery of maternal placental (intervillous) blood for use in immunologic assays is described. 60% of the perfused blood samples tested for fetal red blood cell (RBC) contamination were found to be pure maternal blood; in the remainder, fetal RBC contamination, with a single exception, was less than 6%. The intervillous mononuclear cells (IVBMC) isolated from this blood were of predominantly maternal origin as demonstrated by a polymerase chain reaction-based DNA typing technique. The number of IVBMC obtained was within the range of 9 to 55 X 10(6) cells. Phenotypic analysis of IVBMC surface antigens revealed that 61% of the cells were CD3 + T-cells and 18% were CD19 + B-cells. The CD4 + and CD8 + T-lymphocyte subsets accounted for 28 and 26% of the IVBMC, respectively. The IVBMC were functionally competent as evidenced by in vitro lymphoproliferation and cytokine production in response to mitogen and PPD stimulation. This technique allows for rapid and safe isolation of large numbers of IVBMC which are functionally active up to 12 h post-delivery, thus representing a significant improvement over previously described methods. It should facilitate more vigorous research in the study of uteroplacental immunity and infectious disease research, particularly in field settings where sample collection and laboratory facilities are distant.
Moore
J M
JM
National Center for Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Services, Atlanta, GA 30333, USA.
Nahlen
B
B
Ofulla
A V
AV
Caba
J
J
Ayisi
J
J
Oloo
A
A
Misore
A
A
Nahmias
A J
AJ
Lal
A A
AA
Udhayakumar
V
V
D002824:Chorionic Villi / Q000098:blood supply* / Q000276:immunology / Q000378:metabolism; D016207:Cytokines / Q000096:biosynthesis; D004247:DNA / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005319:Fetal Hemoglobin / Q000032:analysis; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D010477:Perfusion; D010641:Phenotype; D011247:Pregnancy / Q000097:blood* / Q000276:immunology / Q000378:metabolism; D013194:Staining and Labeling
D016207:Cytokines; D004247:DNA; D005319:Fetal Hemoglobin