Methods and compositions relating to assays of fetal extravillous trophoblast cells
AssigneeWayne State University
InventorsD. Randall Armant, Sascha Drewlo
Life of this patent
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Abstract
Methods of isolating and assaying fetal extravillous trophoblast cells, including assays of RNA of the fetal extravillous trophoblast cells according to aspects of the disclosure include obtaining a maternal endocervical sample containing fetal extravillous trophoblast cells from a pregnant subject; fixing the maternal endocervical sample in an aldehyde fixative, removing fetal extravillous trophoblast cells from the maternal endocervical sample thereby producing isolated extravillous trophoblast cells, isolating and assaying RNA from the fetal extravillous trophoblast cells.
Claims17
A method of assaying RNA from fetal extravillous trophoblast cells, comprising:
- obtaining a maternal endocervical sample from a pregnant subject;
- removing fetal extravillous trophoblast cells from the maternal endocervical sample, producing isolated fetal extravillous trophoblast cells;
- fixing fetal extravillous trophoblast cells by treatment with an aldehyde fixative producing aldehyde fixed fetal extravillous trophoblast cells;
- washing the aldehyde fixed fetal extravillous trophoblast cells or washing a lysate of the isolated aldehyde fixed fetal extravillous trophoblast cells to promote removal of the crosslinks introduced by the aldehyde fixative, producing washed fetal extravillous trophoblast cells or washed lysate;
- extracting fetal extravillous trophoblast cell RNA from the washed fetal extravillous trophoblast cells or washed lysate;
- and
- assaying the fetal extravillous trophoblast cell RNA to determine one or more characteristics of the fetal extravillous trophoblast cell RNA.
A method of assaying fetal extravillous trophoblast cells, comprising:
- obtaining a maternal endocervical sample from a pregnant subject;
- contacting the maternal endocervical sample with a first antibody, the first antibody specific for the fetal extravillous trophoblast cells, wherein the first antibody does not bind to maternal cells in the maternal endocervical sample;
- contacting the maternal endocervical sample with one more additional antibodies, wherein the first antibody and each additional antibody is distinguishably labeled with different labels detectable by inductively coupled plasma mass spectrometry;
- nebulizing the maternal endocervical sample to separate cells;
- and
- performing inductively coupled plasma mass spectrometry on the cells, thereby assaying the fetal extravillous trophoblast cells.
Dependencies are inferred from the claim text.