PMID- 17580088
OWN - NLM
STAT- MEDLINE
DCOM- 20071102
LR  - 20121115
IS  - 0093-691X (Print)
IS  - 0093-691X (Linking)
VI  - 68
IP  - 4
DP  - 2007 Sep 1
TI  - Validation of primary epitheloid cell cultures isolated from bovine placental 
      caruncles and cotyledons.
PG  - 592-603
AB  - In order to study feto-maternal interactions in the bovine synepitheliochorial 
      placenta primary cell cultures of both placentomal components throughout 
      pregnancy, namely caruncular epithelial cells and trophoblast cells were 
      developed. The aim of this study was to validate and improve a method to culture 
      caruncular epithelial cells and fetal trophoblast from manually separated 
      placentomes. Prior to seeding the presence of fetal cells in caruncular samples 
      and vice-versa could be demonstrated by the detection of the Y-chromosome via 
      fluorescence in situ hybridization (FISH) provided the fetus was male. Epitheloid 
      shaped cells present in both cultures (cotyledon and caruncle) were characterized 
      on a morphological basis as well as by immunofluorescence and Western blot 
      thereby detecting cytokeratin, zonula occludens-1 and vimentin but not 
      alpha-smooth muscle actin and desmin. The absence of the Y-chromosome 
      demonstrated the caruncular origin of epitheloid cells. In addition, a population 
      of polygonally shaped cells derived from the cotyledon was propagated and 
      displayed the same cytoskeletal characteristics as described above. The presence 
      of the Y-chromosome confirmed the fetal origin of these cells and the lacking 
      uptake of fluorescence conjugated low density lipoprotein, specific for 
      endothelial cells, identified polygonally shaped cells as fetal trophoblast 
      cells. In conclusion, the cross-contamination of maternal and fetal cells in 
      manually separated placentomes should be considered in future experiments as it 
      may lead to false positive results dependent on the sensitivity of the method 
      applied. This study highlights the importance of an appropriate cell 
      characterization and identification, especially when isolating primary cells.
FAU - Bridger, P S
AU  - Bridger PS
AD  - Department of Veterinary Anatomy, Histology and Embryology, 
      Justus-Liebig-University, Frankfurter Str 98, Giessen, Germany.
FAU - Haupt, S
AU  - Haupt S
FAU - Klisch, K
AU  - Klisch K
FAU - Leiser, R
AU  - Leiser R
FAU - Tinneberg, H-R
AU  - Tinneberg HR
FAU - Pfarrer, C
AU  - Pfarrer C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20070618
PL  - United States
TA  - Theriogenology
JT  - Theriogenology
JID - 0421510
RN  - 0 (Membrane Proteins)
RN  - 0 (Phosphoproteins)
RN  - 0 (Vimentin)
RN  - 0 (Zonula Occludens-1 Protein)
RN  - 68238-35-7 (Keratins)
SB  - IM
MH  - Animals
MH  - Blotting, Western/veterinary
MH  - Cattle/*physiology
MH  - Epithelial Cells/*cytology
MH  - Female
MH  - Fluorescent Antibody Technique/veterinary
MH  - In Situ Hybridization, Fluorescence/veterinary
MH  - Keratins/metabolism
MH  - Male
MH  - Membrane Proteins/metabolism
MH  - Phosphoproteins/metabolism
MH  - Placenta/*cytology
MH  - Pregnancy
MH  - Trophoblasts/*cytology
MH  - Vimentin/metabolism
MH  - Y Chromosome
MH  - Zonula Occludens-1 Protein
EDAT- 2007/06/21 09:00
MHDA- 2007/11/06 09:00
CRDT- 2007/06/21 09:00
PHST- 2007/01/22 00:00 [received]
PHST- 2007/04/04 00:00 [revised]
PHST- 2007/05/05 00:00 [accepted]
PHST- 2007/06/21 09:00 [pubmed]
PHST- 2007/11/06 09:00 [medline]
PHST- 2007/06/21 09:00 [entrez]
AID - S0093-691X(07)00264-6 [pii]
AID - 10.1016/j.theriogenology.2007.05.046 [doi]
PST - ppublish
SO  - Theriogenology. 2007 Sep 1;68(4):592-603. doi: 
      10.1016/j.theriogenology.2007.05.046. Epub 2007 Jun 18.