Limits...
Expression of TCR-Vβ peptides by murine bone marrow cells does not identify T-cell progenitors.

Abbey JL, Karsunky H, Serwold T, Papathanasiou P, Weissman IL, O'Neill HC - J. Cell. Mol. Med. (2015)

Bottom Line: No T-cell or myeloid progeny were detected following introduction of cells via the intrathymic or intravenous routes.However, B-cell development was detected in spleen.This pattern of restricted in vivo reconstitution disputes Lin(-) Vβ8.2(+) Cβ(-) BM cells as committed T-cell progenitors, but raises the possibility of progenitors with potential for B-cell development.

View Article: PubMed Central - PubMed

Affiliation: Research School of Biology, Australian National University, Canberra, ACT, Australia.

No MeSH data available.


Related in: MedlinePlus

Analysis of Vβ8+Cβ− cells among bone marrow (BM) progenitors. A population of Lin− BM was prepared from BA (C57BL/Ka-Ly5.1) mice and cells were stained with fluorochrome-conjugated antibodies specific for Vβ8, Cβ, CD127, Sca-1 and c-Kit. FACS analysis was used to distinguish cell subsets. (A) Viable (PI−) lymphoid cells were gated initially on the basis of Side Scatter (SSC) and then Vβ8+Cβ− cells were gated for analysis of marker expression of c-Kit, Sca-1 and CD127 markers to delineate hematopoietic progenitors. Numbers in quadrants reflect % cells staining relative to the initial gated population of viable lymphoid cells. (B) Size comparison of subsets isolated from total lineage depleted (Lin−) BM, resorted Lin−BM and spleen.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4549046&req=5

fig02: Analysis of Vβ8+Cβ− cells among bone marrow (BM) progenitors. A population of Lin− BM was prepared from BA (C57BL/Ka-Ly5.1) mice and cells were stained with fluorochrome-conjugated antibodies specific for Vβ8, Cβ, CD127, Sca-1 and c-Kit. FACS analysis was used to distinguish cell subsets. (A) Viable (PI−) lymphoid cells were gated initially on the basis of Side Scatter (SSC) and then Vβ8+Cβ− cells were gated for analysis of marker expression of c-Kit, Sca-1 and CD127 markers to delineate hematopoietic progenitors. Numbers in quadrants reflect % cells staining relative to the initial gated population of viable lymphoid cells. (B) Size comparison of subsets isolated from total lineage depleted (Lin−) BM, resorted Lin−BM and spleen.

Mentions: Lin− BM cells were prepared and stained with antibodies to Vβ8 and Cβ to gate the Vβ8+Cβ− subset. This population was then analysed for expression of c-Kit, Sca-1 and CD127, and distinct subsets of c-Kit+ hematopoietic progenitors delineated (Fig.2). Clear populations of Sca-1loc-KitloCD127+ (CLP-like) cells, Sca-1−c-KithiCD127− [common myeloid progenitor (CMP)-like], but almost no Sca-1+/hic-Kit+CD127− (HSC- or MPP-like) cells could be distinguished among the Vβ8+Cβ− subset of BM. The Sca-1+c-Kit−CD127− subset does not resemble progenitors and could reflect lymphoid cells. The phenotypes of CLP, CMP, MPP and HSC with respect to these markers have been described previously 15,25,26. A summary of the percentage representation of these subsets among Lin− BM, resorted Lin− BM and spleen is shown in Figure2. While CMP-like and CLP-like cells were present among the Vβ8+Cβ− subset of spleen and BM, the HSC-like or MPP-like subset was less well represented among the BM subset.


Expression of TCR-Vβ peptides by murine bone marrow cells does not identify T-cell progenitors.

Abbey JL, Karsunky H, Serwold T, Papathanasiou P, Weissman IL, O'Neill HC - J. Cell. Mol. Med. (2015)

Analysis of Vβ8+Cβ− cells among bone marrow (BM) progenitors. A population of Lin− BM was prepared from BA (C57BL/Ka-Ly5.1) mice and cells were stained with fluorochrome-conjugated antibodies specific for Vβ8, Cβ, CD127, Sca-1 and c-Kit. FACS analysis was used to distinguish cell subsets. (A) Viable (PI−) lymphoid cells were gated initially on the basis of Side Scatter (SSC) and then Vβ8+Cβ− cells were gated for analysis of marker expression of c-Kit, Sca-1 and CD127 markers to delineate hematopoietic progenitors. Numbers in quadrants reflect % cells staining relative to the initial gated population of viable lymphoid cells. (B) Size comparison of subsets isolated from total lineage depleted (Lin−) BM, resorted Lin−BM and spleen.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4549046&req=5

fig02: Analysis of Vβ8+Cβ− cells among bone marrow (BM) progenitors. A population of Lin− BM was prepared from BA (C57BL/Ka-Ly5.1) mice and cells were stained with fluorochrome-conjugated antibodies specific for Vβ8, Cβ, CD127, Sca-1 and c-Kit. FACS analysis was used to distinguish cell subsets. (A) Viable (PI−) lymphoid cells were gated initially on the basis of Side Scatter (SSC) and then Vβ8+Cβ− cells were gated for analysis of marker expression of c-Kit, Sca-1 and CD127 markers to delineate hematopoietic progenitors. Numbers in quadrants reflect % cells staining relative to the initial gated population of viable lymphoid cells. (B) Size comparison of subsets isolated from total lineage depleted (Lin−) BM, resorted Lin−BM and spleen.
Mentions: Lin− BM cells were prepared and stained with antibodies to Vβ8 and Cβ to gate the Vβ8+Cβ− subset. This population was then analysed for expression of c-Kit, Sca-1 and CD127, and distinct subsets of c-Kit+ hematopoietic progenitors delineated (Fig.2). Clear populations of Sca-1loc-KitloCD127+ (CLP-like) cells, Sca-1−c-KithiCD127− [common myeloid progenitor (CMP)-like], but almost no Sca-1+/hic-Kit+CD127− (HSC- or MPP-like) cells could be distinguished among the Vβ8+Cβ− subset of BM. The Sca-1+c-Kit−CD127− subset does not resemble progenitors and could reflect lymphoid cells. The phenotypes of CLP, CMP, MPP and HSC with respect to these markers have been described previously 15,25,26. A summary of the percentage representation of these subsets among Lin− BM, resorted Lin− BM and spleen is shown in Figure2. While CMP-like and CLP-like cells were present among the Vβ8+Cβ− subset of spleen and BM, the HSC-like or MPP-like subset was less well represented among the BM subset.

Bottom Line: No T-cell or myeloid progeny were detected following introduction of cells via the intrathymic or intravenous routes.However, B-cell development was detected in spleen.This pattern of restricted in vivo reconstitution disputes Lin(-) Vβ8.2(+) Cβ(-) BM cells as committed T-cell progenitors, but raises the possibility of progenitors with potential for B-cell development.

View Article: PubMed Central - PubMed

Affiliation: Research School of Biology, Australian National University, Canberra, ACT, Australia.

No MeSH data available.


Related in: MedlinePlus