27-04-2017 дата публикации
Номер: US20170115282A1
Принадлежит:
The invention is in the field of analysis of cell populations and cell separation and the compositions obtained thereby. More particularly, the invention concerns a method for separating cells of interest according to a specific marker expressed on their surface. 1. A method for separating spermatozoa according to the specific marker CD52 (Human epididymal secreted protein) expressed on their surface from a plurality of cells contained , in a dry state , on an inert substrate , which method comprises the steps ofi) obtaining the plurality of cells contained on the inert substrate,ii) contacting said plurality of cells with a serum-free buffer comprising Phosphate buffered saline 1× and about 2-20 mM of EDTA, at about 10-30° C. during 10-45 min and exposing the plurality of cells to at least one magnetic particle under conditions effective to allow direct or indirect coupling of the magnetic particle to the specific marker CD52 expressed on the surface of the spermatozoa,iii) recovering the plurality of cells, andiv) separating the spermatozoa that are coupled to the magnetic particle from the cells not coupled to the magnetic particle.2. The method of claim 1 , wherein the serum-free buffer further comprises about 10-200 ng/ml of proteinase K.3. The method of claim 1 , wherein the serum-free buffer comprises about 2-10 mM of EDTA.4. The method of claim 1 , wherein said plurality of cells is contacted with the serum-free buffer at 25° C. during 30 min.5. The method of claim 1 , wherein the cells of the plurality of cells contained on an inert substrate are selected from the group consisting of spermatozoa claim 1 , epithelial cells claim 1 , vaginal cells claim 1 , buccal epithelial cells claim 1 , white blood cells and red blood cells.6. The method of claim 1 , wherein the method further comprises a step of washing the plurality of cells contacted with the serum-free buffer with a washing buffer before exposing said plurality of cells to the at least one magnetic ...
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