title: Proteomic analysis of human blastocoel fluid and blastocyst cells.
abstract: Human embryonic stem cells (hESCs) are derived from the inner cell mass (ICM) of the blastocyst and can differentiate into any cell type in the human body. These cells hold a great potential for regenerative medicine, but to obtain enough cells needed for medical treatment, culture is required on a large scale. In the undifferentiated state, hESCs appear to possess an unlimited potential for proliferation, but optimal, defined, and safe culture conditions remain a challenge. The aim of the present study was to identify proteins in the natural environment of undifferentiated hESCs, namely, the blastocoel fluid, which is in contact with all the cells in the blastocyst, including hESCs. Fifty-three surplus human blastocysts were donated after informed consent, and blastocoel fluid was isolated by micromanipulation. Using highly sensitive nano-high-pressure liquid chromatography-tandem mass spectrometry, 286 proteins were identified in the blastocoel fluid and 1,307 proteins in the corresponding cells of the blastocyst. Forty-two were previously uncharacterized proteins-8 of these originated from the blastocoel fluid. Furthermore, several heat shock proteins (Hsp27, Hsp60, Hsc70, and Hsp90) were identified in blastocoel fluid together with zona pellucida proteins (ZP2-4), Vitamin D-binding protein, and Retinol-binding protein 4. Proteins that regulate ciliary assembly and function were also identified, including Bardet-Biedl syndrome protein 7. This study has identified numerous proteins that cells from the ICM of the human blastocyst are exposed to via the blastocoel fluid. These results can be an inspiration for the development of improved culture conditions for hESCs.
authors.affiliation: Laboratory of Reproductive Biology, University Hospital of Copenhagen, Copenhagen, Denmark. pljensen@origio.com
authors.forename: Pernille Linnert
authors.initials: PL
authors.lastname: Jensen
authors.forename: Hans Christian
authors.initials: HC
authors.lastname: Beck
authors.forename: Jørgen
authors.initials: J
authors.lastname: Petersen
authors.forename: Julius
authors.initials: J
authors.lastname: Hreinsson
authors.forename: Kjell
authors.initials: K
authors.lastname: Wånggren
authors.forename: Steen B
authors.initials: SB
authors.lastname: Laursen
authors.forename: Pernille Dissing
authors.initials: PD
authors.lastname: Sørensen
authors.forename: Søren Tvorup
authors.initials: ST
authors.lastname: Christensen
authors.forename: Claus Yding
authors.initials: CY
authors.lastname: Andersen
mesh_terms: D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D046149:Embryo Culture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D014809:Vitamin D-Binding Protein / Q000378:metabolism; D015044:Zona Pellucida / Q000378:metabolism
chemical_list: D020543:Proteome; D014809:Vitamin D-Binding Protein