Human decidual production of hepatocyte-growth factor is not influenced by trophoblastic invasion in vivo.

OBJECTIVE: To evaluate and compare production of hepatocyte growth factor (HGF) from human first-trimester implantation-site decidua (decidua basalis) and nonimplantation site decidua (decidua parietalis), and hence to determine whether human trophoblast invasion in vivo is associated with increased decidual HGF production. DESIGN: Controlled prospective study. SETTING: University hospital-based study. PATIENT(S): Ten women undergoing first-trimester termination of singleton pregnancy for psychosocial reasons without preexisting medical or gynecologic diseases. INTERVENTION(S): Decidual samples surgically excised and processed for paraffin-embedded immunohistochemistry and for reverse transcription-polymerase chain reaction (RT-PCR) studies. MAIN OUTCOME MEASURE(S): Protein and mRNA production in decidua basalis and decidua parietalis by immunohistochemistry and RT-PCR, respectively. RESULT(S): No statistically significant difference was found between decidua basalis and decidua parietalis in HGF protein or mRNA production. Immunohistochemical analysis (n = 9) showed a mean score of 3.28 +/- 2.37 for decidua basalis and 3.61 +/- 2.66 for decidua parietalis. Semiquantitative analysis of HGF mRNA expression between the two sites showed no statistically significant difference (n = 10) CONCLUSION(S): Human decidual production of HGF is not influenced by trophoblastic invasion in vivo.

Corticotropin-releasing hormone peptide and human first-trimester placental growth.

The aim of this study was to determine whether corticotropin-releasing hormone (CRH) regulates human trophoblast cell growth. The results showed that exogenous CRH significantly stimulated human trophoblast proliferation in first-trimester primary cultures. In vivo, CRH was strongly immunolocalised to cytotrophoblastic cells in proliferative cell columns and in chorionic villi. We postulate that CRH may have an important role in early placental development and successful pregnancy.

Lack of site-specific production of decidual alpha-2 macroglobulin in human pregnancy.

BACKGROUND: Alpha-2 Macroglobulin (A2M) is a protease inhibitor that is present in both human and rat decidual tissue. In mice, decidual A2M prevents excessive trophoblastic invasion; however, its role in human decidual tissue is unknown. It is possible that A2M may also influence trophoblast invasion in human pregnancy, which would be reflected in increased A2M production in decidua basalis. The aim of the current study was to determine and compare A2M production from first trimester human decidua basalis and decidua parietalis. METHODS: Human decidual tissues were obtained from patients undergoing surgical termination at 9 to 12 gestational weeks. Strips of decidua basalis and decidua parietalis were obtained by uterine curettage under real-time ultrasound guidance. Tissue samples were fixed in 10% formalin or snap-frozen for immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, respectively. Protein and mRNA production between the two sites were compared using the Mann-Whitney U test. RESULTS: Paired basal and parietal decidua were analyzed by immunohistochemistry (n = 9) and by RT-PCR (n = 10). There was no significant difference in A2M mRNA expression between decidua basalis and decidua parietalis (P = .5). Immunohistochemical staining intensity for A2M protein was significantly higher in basalis than in parietalis (P = .004), but the extent of positively stained cells were not significantly different (P = .051). Strong A2M staining in decidua basalis was mainly localized in the intracellular storage vesicles, which may suggest a role of A2M in this site. CONCLUSIONS: We conclude that the expression pattern of A2M in human decidua basalis and decidua parietalis is not consistent with an important role of this gene during the observed gestational period. Contrary to its role in rodent implantation, A2M is probably not involved in regulating human implantation and trophoblastic invasion during this gestational window frame.

Establishment and characterization of a human first-trimester extravillous trophoblast cell line (TEV-1).

OBJECTIVE: Research into the biology of human trophoblast invasion has been hampered by a lack of in vitro models. The aim of this study was to establish and characterize a human extravillous trophoblast cell line from the first-trimester placenta. METHODS: Human papillomavirus type 16 (HPV16) E6/E7 genes were stably expressed in primary cultures of first-trimester placenta via a retroviral vector (pLXSN-E6/E7). Several clones were characterized for extravillous trophoblastic properties by reverse transcriptase polymerase chain reaction (RT-PCR), Western blotting, and immunocytochemistry. The activities of matrix metalloproteinase (MMP)-2 and MMP-9 were examined with gelatin zymography. One clone (TEV-1), which retains all the established criteria for extravillous trophoblasts, was used in microarray analysis with Stanford Human cDNA chip (41, 421 cDNA features) to examine the differential gene expression after treatment of transforming growth factor beta 1 (TGFbeta1). The responsive gene to TGFbeta1 treatment was confirmed by quantitative real-time PCR. RESULTS: The clonal TEV-1 has been passaged for more than 105 population doublings with no sign of senescence, the activation of telomerase at early passages, and a near-diploid karyotype. TEV-1 cells expressed cytokeratin 7, HLA-G (a histocompatibility antigen, class IB), and CD9 (the cluster of differentiation antigen 9), and secreted active MMP-2 and MMP-9. TGFbeta1 treatment altered the gene expression profile of TEV-1 cells with a marked up-regulation of insulin-like growth factor binding protein 3 (IGFBP3), which was confirmed by quantitative real-time PCR. In addition, the TEV-1 was nontumorigenic when injected into nude mice and unable to form colonies in soft agar. CONCLUSION: Phenotypic and biologic characteristics of TEV-1 were shown as the properties of extravillous trophoblasts; thus, the TEV-1 cell line may be used as a cell model in extravillous trophoblast studies.