Index sorting resolves heterogeneous murine hematopoietic stem cell populations

R Schulte; NK Wilson; JCM Prick; C Cossetti; MK Maj; B Gottgens; DG Kent

doi:10.1016/j.exphem.2015.05.006

Index sorting resolves heterogeneous murine hematopoietic stem cell populations

R Schulte, NK Wilson, JCM Prick, C Cossetti, MK Maj, B Gottgens, DG Kent

Biology

Research output: Contribution to journal › Article › peer-review

Abstract

Recent advances in the cellular and molecular biology of single stem cells have uncovered significant heterogeneity in the functional properties of stem cell populations. This has prompted the development of approaches to study single cells in isolation, often performed using multiparameter flow cytometry. However, many stem cell populations are too rare to test all possible cell surface marker combinations, and virtually nothing is known about functional differences associated with varying intensities of such markers. Here we describe the use of index sorting for further resolution of the flow cytometric isolation of single murine hematopoietic stem cells (HSCs). Specifically, we associate single-cell functional assay outcomes with distinct cell surface marker expression intensities. High levels of both CD150 and EPCR associate with delayed kinetics of cell division and low levels of differentiation. Moreover, cells that do not form single HSC-derived clones appear in the 7AAD(dim) fraction, suggesting that even low levels of 7AAD staining are indicative of less healthy cell populations. These data indicate that when used in combination with single-cell functional assays, index sorting is a powerful tool for refining cell isolation strategies. This approach can be broadly applied to other single-cell systems, both to improve isolation and to acquire additional cell surface marker information.

Original language	English
Pages (from-to)	803-811
Number of pages	9
Journal	Experimental hematology
Volume	43
Issue number	9
DOIs	https://doi.org/10.1016/j.exphem.2015.05.006
Publication status	Published - 1 Sep 2015

Access to Document

10.1016/j.exphem.2015.05.006

https://www.ncbi.nlm.nih.gov/pubmed/26051918

Cite this

@article{225bfaab0291457fbb586777da83e487,

title = "Index sorting resolves heterogeneous murine hematopoietic stem cell populations",

abstract = "Recent advances in the cellular and molecular biology of single stem cells have uncovered significant heterogeneity in the functional properties of stem cell populations. This has prompted the development of approaches to study single cells in isolation, often performed using multiparameter flow cytometry. However, many stem cell populations are too rare to test all possible cell surface marker combinations, and virtually nothing is known about functional differences associated with varying intensities of such markers. Here we describe the use of index sorting for further resolution of the flow cytometric isolation of single murine hematopoietic stem cells (HSCs). Specifically, we associate single-cell functional assay outcomes with distinct cell surface marker expression intensities. High levels of both CD150 and EPCR associate with delayed kinetics of cell division and low levels of differentiation. Moreover, cells that do not form single HSC-derived clones appear in the 7AAD(dim) fraction, suggesting that even low levels of 7AAD staining are indicative of less healthy cell populations. These data indicate that when used in combination with single-cell functional assays, index sorting is a powerful tool for refining cell isolation strategies. This approach can be broadly applied to other single-cell systems, both to improve isolation and to acquire additional cell surface marker information.",

author = "R Schulte and NK Wilson and JCM Prick and C Cossetti and MK Maj and B Gottgens and DG Kent",

year = "2015",

month = sep,

day = "1",

doi = "10.1016/j.exphem.2015.05.006",

language = "English",

volume = "43",

pages = "803--811",

journal = "Experimental Haematology",

issn = "0301-472X",

publisher = "Elsevier",

number = "9",

}

TY - JOUR

T1 - Index sorting resolves heterogeneous murine hematopoietic stem cell populations

AU - Schulte, R

AU - Wilson, NK

AU - Prick, JCM

AU - Cossetti, C

AU - Maj, MK

AU - Gottgens, B

AU - Kent, DG

PY - 2015/9/1

Y1 - 2015/9/1

N2 - Recent advances in the cellular and molecular biology of single stem cells have uncovered significant heterogeneity in the functional properties of stem cell populations. This has prompted the development of approaches to study single cells in isolation, often performed using multiparameter flow cytometry. However, many stem cell populations are too rare to test all possible cell surface marker combinations, and virtually nothing is known about functional differences associated with varying intensities of such markers. Here we describe the use of index sorting for further resolution of the flow cytometric isolation of single murine hematopoietic stem cells (HSCs). Specifically, we associate single-cell functional assay outcomes with distinct cell surface marker expression intensities. High levels of both CD150 and EPCR associate with delayed kinetics of cell division and low levels of differentiation. Moreover, cells that do not form single HSC-derived clones appear in the 7AAD(dim) fraction, suggesting that even low levels of 7AAD staining are indicative of less healthy cell populations. These data indicate that when used in combination with single-cell functional assays, index sorting is a powerful tool for refining cell isolation strategies. This approach can be broadly applied to other single-cell systems, both to improve isolation and to acquire additional cell surface marker information.

AB - Recent advances in the cellular and molecular biology of single stem cells have uncovered significant heterogeneity in the functional properties of stem cell populations. This has prompted the development of approaches to study single cells in isolation, often performed using multiparameter flow cytometry. However, many stem cell populations are too rare to test all possible cell surface marker combinations, and virtually nothing is known about functional differences associated with varying intensities of such markers. Here we describe the use of index sorting for further resolution of the flow cytometric isolation of single murine hematopoietic stem cells (HSCs). Specifically, we associate single-cell functional assay outcomes with distinct cell surface marker expression intensities. High levels of both CD150 and EPCR associate with delayed kinetics of cell division and low levels of differentiation. Moreover, cells that do not form single HSC-derived clones appear in the 7AAD(dim) fraction, suggesting that even low levels of 7AAD staining are indicative of less healthy cell populations. These data indicate that when used in combination with single-cell functional assays, index sorting is a powerful tool for refining cell isolation strategies. This approach can be broadly applied to other single-cell systems, both to improve isolation and to acquire additional cell surface marker information.

U2 - 10.1016/j.exphem.2015.05.006

DO - 10.1016/j.exphem.2015.05.006

M3 - Article

VL - 43

SP - 803

EP - 811

JO - Experimental Haematology

JF - Experimental Haematology

SN - 0301-472X

IS - 9

ER -