Secretory characteristics and viability of human term placental tissue after overnight cold preservation.
Collection of human term placentae for research purposes is generally limited during working hours. Preserving placental tissue overnight might help to postpone experiments and, by extent, to increase material availability. In this study, fragments from normal placentae were incubated at 37 degrees C either immediately after delivery or after preservation at 4 degrees C in a HEPES-buffered solution or in a Roswell Park Memorial Institute (RPMI) 1640 culture medium. Protein, human chorionic gonadotrophin (HCG), human placental lactogen (HPL) and lactate dehydrogenase (LDH) contents within preserved explants were similar to those within freshly delivered ones. In contrast, HCG and HPL amounts released during incubation of preserved tissue were lower than with freshly delivered tissue. Differences were significant only during the first 3 h of incubation. Hormone releases were similarly Ca(2+)-stimulated, and Co(2+)- and low temperature-inhibited in preserved and freshly delivered tissues. After preservation, LDH leakage was also reduced. Furthermore, before and after 37 degrees C incubation during 6 h, preserved tissue was morphologically indistinguishable from freshly delivered tissue and showed neither higher incidence of DNA fragmentation, nor elevated caspase-3 activity, both of which are markers of apoptosis. This study validates an original, useful and rapid method to preserve placental tissue. Consequently, this preservation model may facilitate the study of physiological processes regulating placental hormone secretion in normal and pathological conditions.
Cirelli
N
N
Research Laboratory on Reproduction, Université Libre de Bruxelles (ULB), Brussels, Belgium.
Lebrun
P
P
Gueuning
C
C
Moens
A
A
Delogne-Desnoeck
J
J
Dictus-Vermeulen
C
C
Vanbellinghen
A
A
Meuris
S
S
D017209:Apoptosis; D002021:Buffers; D002118:Calcium / Q000494:pharmacology; D053148:Caspase 3; D020169:Caspases / Q000032:analysis; D006063:Chorionic Gonadotropin / Q000032:analysis / Q000378:metabolism; D003035:Cobalt / Q000494:pharmacology; D003080:Cold Temperature; D003470:Culture Media; D053938:DNA Fragmentation; D005260:Female; D006801:Humans; D020287:In Situ Nick-End Labeling; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000378:metabolism; D010920:Placenta / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D010928:Placental Lactogen / Q000032:analysis / Q000378:metabolism; D011247:Pregnancy; D011506:Proteins / Q000032:analysis; D013997:Time Factors; D014021:Tissue Preservation
D002021:Buffers; D006063:Chorionic Gonadotropin; D003470:Culture Media; D011506:Proteins; D003035:Cobalt; D010928:Placental Lactogen; D007770:L-Lactate Dehydrogenase; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D002118:Calcium