Neutrophil progenitors and related methods and uses
There is provided a method of identifying a neutrophil progenitor, the method comprising: determining an expression of at least one biomarker selected from the group consisting of: CD71, LOX-1, CD164, CD112, CD181, TACSTD2, CD11b and CD49d in a cell. In various embodiments, the cell is identified as a neutrophil progenitor when it is determined to have at least one of the following expression profiles: CD71 hi/+ , LOX-1 int/lo/− , CD164 hi/+ , CD112 hi/+ , CD181 int/lo/− , TACSTD2 hi/+ , CD11b lo/− and/or CD49d int/hi/+ . Also disclosed are a method of sorting and/or separating neutrophil progenitors from a cell population, a composition that is enriched in neutrophil progenitors and related uses and methods.
1 . A method of identifying neutrophil progenitors, the method comprising:
determining an expression of the following biomarkers: CD71, LOX-1, CD164, CD112, CD181, TACSTD2, CD11b and CD49d in a cell;
identifying the cell as an early neutrophil progenitor when it is determined to have the following expression profiles: CD71 hi/+ , LOX-1 int/lo/− , CD164 hi/+ , CD112 hi/+ , CD181 int/lo/− TACSTD2 hi/+ , CD11b lo/− and CD49d int/hi/+ ;
selecting the cell identified as the early neutrophil progenitor;
determining an expression of the selected cell of a further biomarker CD49d and a side-scatter (SSC) property of the neutrophil progenitor;
identifying the cell as an early committed neutrophil progenitor or an intermediate neutrophil progenitor subtype based on the expression of the further biomarker and the side-scatter property,
wherein where the cell is determined to be CD71 hi/+ , CD49d hi/+ and SSC lo , the cell is identified as an early committed neutrophil progenitor, and
wherein where the cell is determined to be CD71 hi/+ , CD49d int/lo/− and SSC hi , the cell is identified as an intermediate neutrophil progenitor that is downstream in neutrophil lineage to the early committed neutrophil progenitor,
wherein the method further comprising culturing the identified neutrophil progenitors to obtain proliferation and/or differentiation of the neutrophil progenitors to obtain progenies thereof, and
administering the neutrophil progenitors and/or the progenies thereof to a subject in need thereof.
2 . The method according to claim 1 , wherein determination of the expression of the biomarkers and the further biomarker comprises contacting the cell with antibodies against the biomarkers and the further biomarker.
3 . A method of sorting and/or separating neutrophil progenitors from a cell population, the method comprising:
selecting the cell having the following expression profiles: CD71 hi/+ , LOX-1 int/lo/− CD164 hi/+ , CD112 hi/+ , CD181 int/lo/− TACSTD2 hi/+ , CD11b lo/− and CD49d int/hi/+ ;
determining an expression of the selected cells of a further biomarker CD49d and a side-scatter (SSC) property;
identifying the cells as an early committed neutrophil progenitor or an intermediate neutrophil progenitor subtype based on the expression of the further biomarker and the side-scatter property;
sorting and/or separating the identified cells as an early committed neutrophil progenitor having the following expression profiles: CD71 hi/+ , CD49d hi/+ and SSC lo ; and
sorting and/or separating the identified cells as an intermediate neutrophil progenitor having the following expression profiles: CD71 hi/+ , CD49d int/lo/− and SSC hi .
4 . The method according to claim 3 , wherein the cell population is derived from cord blood and/or bone marrow.
5 . The method according to claim 3 , the method further comprising culturing the neutrophil progenitors to obtain proliferation and/or differentiation of the neutrophil progenitors to obtain progenies thereof.
6 . The method according to claim 3 , the method further comprising administering the neutrophil progenitors and/or the progenies thereof to a subject in need thereof.
7 . The method according to claim 6 , wherein the subject has neutropenia.
8 . The method according to claim 3 , wherein the selecting comprises contacting the cells with antibodies against CD71, LOX-1, CD164, CD112, CD181, TACSTD2, CD11b and CD49d.
9 . A composition that is enriched in early neutrophil progenitors having the following expression profiles: CD71 hi/+ , CD49d hi/+ , SSC lo , LOX-1 int/lo/− , CD164 hi/+ , CD112 hi/+ , CD181 int/lo/− , TACSTD2 hi/+ , and CD11b lo/− .
10 . The method of claim 3 , further comprising preparing a transfusion composition, wherein the composition is an enriched composition of early neutrophil progenitors having CD71 hi/+ CD49d hi+ , SSC lo LOX-1 int/lo/− , CD164 hi/+ , CD112 hi/+ , CD181 int/lo/− , TACSTD2 hi/+ , and CD11b lo/− .
11 . The composition of claim 9 , further comprising an intermediate neutrophil progenitor that is downstream in neutrophil lineage to the early committed neutrophil progenitor having the following expression profile: CD71 hi/30 , CD49d int/lo/− and SSC hi .