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Isolation, purification and in vitro differentiation of cytotrophoblast cells from human term placenta

Overview of attention for article published in Reproductive Biology and Endocrinology, July 2015
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Title
Isolation, purification and in vitro differentiation of cytotrophoblast cells from human term placenta
Published in
Reproductive Biology and Endocrinology, July 2015
DOI 10.1186/s12958-015-0070-8
Pubmed ID
Authors

Liping Li, Danny J Schust

Abstract

The syncytialization of cytotrophoblast cells to syncytiotrophoblast is central to human placental transport and hormone production. Many techniques for in vitro study of this process have been proposed and new investigators to the field may find the literature in the field daunting. Here, we present a straightforward and reliable method to establish this important model using modern but readily available tools and reagents. Villous cytotrophoblast cells are obtained from term placenta using mild enzymatic degradation, Percoll gradient centrifugation, negative magnetic cell sorting using an antibody against classical major histocompatibility complex molecules and in vitro culture on a matrix-coated growth surface. The purity of isolated cytotrophoblast cells exceeds 98 % as assessed by cytokeratin-7 expression using flow cytometry. Contamination by mesenchymal cells, extravillous trophoblast cells, leukocytes, Hofbauer and endothelial cells is minimized (less than 2 % when analyzed for vimentin, HLA-G, CD45, CD163 and CD31 using flow cytometry). Isolated cytotrophoblast cells began to aggregate into monolayers of mononucleated cells within about 12 h of plating. By 72 h in culture, most cytotrophoblast cells have differentiated into syncytiotrophoblast as demonstrated by a loss of intercellular E-cadherin expression upon fusion into multinucleated syncytia. After 72 h in culture, nearly every cultured cell expresses syncytiotrophoblast markers, including cytokeratin-7, human chorionic gonadotropin-β (β-hCG) and the fusion-related proteins glial cell missing-1 (GCM-1) and syncytin. We present an efficient and reliable method for isolating of cytotrophoblast cells with high purity and complete differentiation into syncytiotrophoblast in vitro.

Twitter Demographics

The data shown below were collected from the profile of 1 tweeter who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 121 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United States 1 <1%
Unknown 120 99%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 31 26%
Student > Master 16 13%
Researcher 16 13%
Student > Bachelor 11 9%
Student > Doctoral Student 9 7%
Other 19 16%
Unknown 19 16%
Readers by discipline Count As %
Agricultural and Biological Sciences 25 21%
Biochemistry, Genetics and Molecular Biology 24 20%
Medicine and Dentistry 16 13%
Immunology and Microbiology 12 10%
Pharmacology, Toxicology and Pharmaceutical Science 6 5%
Other 11 9%
Unknown 27 22%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 22 July 2015.
All research outputs
#9,963,015
of 12,445,189 outputs
Outputs from Reproductive Biology and Endocrinology
#341
of 498 outputs
Outputs of similar age
#162,721
of 236,148 outputs
Outputs of similar age from Reproductive Biology and Endocrinology
#3
of 4 outputs
Altmetric has tracked 12,445,189 research outputs across all sources so far. This one is in the 11th percentile – i.e., 11% of other outputs scored the same or lower than it.
So far Altmetric has tracked 498 research outputs from this source. They typically receive more attention than average, with a mean Attention Score of 7.6. This one is in the 20th percentile – i.e., 20% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 236,148 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 17th percentile – i.e., 17% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 4 others from the same source and published within six weeks on either side of this one.