Two-sided culture of human placental trophoblast. Morphology, immunocytochemistry and permeability properties

Bullen, Bridget E.; Bloxam, David L.; Ryder, Timothy A.; Mobberley, Margaret A.; Bax, Bridget E.

doi:10.1016/s0143-4004(05)80217-6

Cited by 29 publications

(11 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus far, PLAP served as a presumptive marker for STBs for the morphological characterization of these cultures. In line with this hypothesis, freshly isolated trophoblasts were found to be cytokeratin-positive (Bax et al 1989;Bullen et al 1990), but only 10% of this population were PLAP-positive cells (i.e., CTBs); after 48 hr in culture all cells expressed PLAP (Bullen et al 1990). However, this is in contrast to the results presented in this investigation, in which almost all isolated cytokeratin-positive cells ( ϭ trophoblasts) were also PLAP-positive immediately after isolation (compare Figures 2A and 2B).…”

Section: Figurementioning

confidence: 69%

Placental Alkaline Phosphatase Expression at the Apical and Basal Plasma Membrane in Term Villous Trophoblasts

Leitner

Szlauer

Ellinger

et al. 2001

J Histochem Cytochem.

View full text Add to dashboard Cite

S U M M A R YHuman placental alkaline phosphatase (PLAP) was localized at the apical and basal plasma membrane of syncytiotrophoblasts and at the surface of cytotrophoblasts in term chorionic villi using immunoelectron microscopy. Similarly, apical and basolateral PLAP expression was found in polarized trophoblast-derived BeWo cells. Trophoblasts isolated from term placentas exhibited mainly vesicular PLAP immunofluorescence staining immediately after isolation. After in vitro differentiation into syncytia, PLAP plasma membrane expression was upregulated and exceeded that observed in mononuclear trophoblasts. These data call for caution in using PLAP as a morphological marker to differentiate syncytiotrophoblasts from cytotrophoblasts or as a marker enzyme for placental brush-border membranes.

show abstract

Section: Figurementioning

confidence: 69%

Placental Alkaline Phosphatase Expression at the Apical and Basal Plasma Membrane in Term Villous Trophoblasts

Leitner

Szlauer

Ellinger

et al. 2001

J Histochem Cytochem.

View full text Add to dashboard Cite

show abstract

“…Previously, the in vitro permeability of cell free amnion was reported to be 1.5 × 10 -6 cm 2 /s for D-glucose and 2-aminoisobutyrate[53]. Further, the in vitro permeability across chorion for meperidine (Mw = 247.33 Da) and diazepam (Mw = 284.7 Da) was reported to be 5.26 × 10 -6 and 4.51 × 10 -6 cm 2 /s respectively.…”

Section: Resultsmentioning

confidence: 99%

Transport and biodistribution of dendrimers across human fetal membranes: Implications for intravaginal administration of dendrimer-drug conjugates

et al. 2010

View full text Add to dashboard Cite

Dendrimers are emerging as promising topical antimicrobial agents, and as targeted nanoscale drug delivery vehicles. Topical intravaginal antimicrobial agents are prescribed to treat the ascending genital infections in pregnant women. The fetal membranes separate the extra-amniotic space and fetus. The purpose of the study is to determine if the dendrimers can be selectively used for local intravaginal application to pregnant women without crossing the membranes into the fetus. In the present study, the transport and permeability of PAMAM (poly(amidoamine)) dendrimers, across human fetal membrane (using a side-by-side diffusion chamber), and its biodistribution (using immunofluorescence) are evaluated ex-vivo. Transport across human fetal membranes (from the maternal side) was evaluated using Fluorescein (FITC), an established transplacental marker (positive control, size~ 400 Da) and fluorophore-tagged G4-PAMAM dendrimers (~ 16 kDa). The fluorophore-tagged G4-PAMAM dendrimers were synthesized and characterized using 1H NMR, MALDI TOF-MS and HPLC analysis. Transfer was measured across the intact fetal membrane (chorioamnion), and the separated chorion and amnion layers. Over a five hour period, the dendrimer transport across all the three membranes was less than < 3 %, whereas the transport of FITC was relatively fast with as much as 49% transport across the amnion. The permeability of FITC (7.9 × 10-7 cm2/s) through the chorioamnion was 7-fold higher than that of the dendrimer (5.8 × 10-8 cm2/s). The biodistribution showed that the dendrimers were largely present in interstitial spaces in the decidual stromal cells and the chorionic trophoblast cells (in 2.5 to 4 h) and surprisingly, to a smaller extent internalized in nuclei of trophoblast cells and nuclei and cytoplasm of stromal cells. Passive diffusion and paracellular transport appear to be the major route for dendrimer transport. The overall findings further suggest that entry of drugs conjugated to dendrimers would be restricted across the human fetal membranes when administered topically by intravaginal route, suggesting new ways of selectively delivering therapeutics to the mother without affecting the fetus.

show abstract

“…We have confirmed that molecular markers ranging from 2-aminoisobutyrate (relative molecular mass (Mr) 103) to human serum albumin (Mr 68 000) readily cross the amniotic membrane support by passive diffusion and that the cell layer forms a significant barrier after 2 d of culture. However, not surprisingly, the clearance of extracellular markers in the presence of the cell layer was still appreciable indicating a measurable leak (Bullen et al 1990).…”

Section: Morphology and Immunocytochemistry The Morphology And Immumentioning

confidence: 97%

“…This method, which is described in detail elsewhere (Bax et al 1989;Bullen et al 1990), removes the need for a purification step. These cells were cultured in supplemented medium 199, buffered at pH 7.4 with either carbon dioxide-air (5:95, v/v), or 20 mmol HEPESA (Bax et al 1989).…”

Section: Isolation Of Trophoblast Cellsmentioning

confidence: 99%

See 1 more Smart Citation

Human placental trophoblast culture: One-sided and two-sided models

Bloxam

1991

proc nutr soc

Self Cite

View full text Add to dashboard Cite

Two-sided culture of human placental trophoblast. Morphology, immunocytochemistry and permeability properties

Cited by 29 publications

References 27 publications

Placental Alkaline Phosphatase Expression at the Apical and Basal Plasma Membrane in Term Villous Trophoblasts

Placental Alkaline Phosphatase Expression at the Apical and Basal Plasma Membrane in Term Villous Trophoblasts

Transport and biodistribution of dendrimers across human fetal membranes: Implications for intravaginal administration of dendrimer-drug conjugates

Human placental trophoblast culture: One-sided and two-sided models

Contact Info

Product

Resources

About