Evaluation of the placental environment with a new in vitro model of histocultures of early and term placentae: determination of cytokine and chemokine expression profiles.
We aimed to set up and validate a new in vitro model of placental histocultures, for the evaluation of cytokine and chemokine profiles of the placental environment, over a long culture period. Micro-explant cultures from 6 early and 6 term placentae were set up on collagen sponge gel supports at a liquid/air interface. At various times during culture, we analyzed tissue morphology and cell death by microscopy and quantified beta-hCG production and mRNA levels for beta-hCG and insulin-like 4 (INSL4). Levels of IL-6, LIF, TNF alpha, IL-10, IFN-gamma, IL-16 and RANTES in the medium were measured by ELISA on days 1, 4 and 7 of culture. SDF-1 mRNA expression was determined by real-time PCR at the same time points. Histocultures from early and term placentae remained viable until day 10. High levels of IL-6 and LIF production, low levels of TNF alpha, IL-10 and IFN-gamma production and significant SDF-1 expression were observed. These data indicate that placental histoculture is a suitable and reliable in vitro model for studying the placental environment.
Faye
A
A
Biology of Retroviruses Unit, Institut Pasteur, 28 rue du Dr Roux, 75015 Paris, France.
Pornprasert
S
S
Dolcini
G
G
Ave
P
P
Taïeb
J
J
Taupin
J L
JL
Derrien
M
M
Huerre
M
M
Barré-Sinoussi
F
F
Chaouat
G
G
Menu
E
E
D000328:Adult; D017209:Apoptosis; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D054377:Chemokine CXCL12; D018925:Chemokines / Q000032:analysis / Q000378:metabolism*; D019743:Chemokines, CXC / Q000235:genetics / Q000378:metabolism; D002824:Chorionic Villi / Q000033:anatomy & histology / Q000378:metabolism* / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D020287:In Situ Nick-End Labeling; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D047929:Term Birth
C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D018925:Chemokines; D019743:Chemokines, CXC; D012333:RNA, Messenger