Colony formation of clone-sorted human hematopoietic progenitors

Hideo Ema, Toshio Suda, Yasusada Miura, Hiromitsu Nakauchi

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

To characterize human hematopoietic progenitors, we performed methylcellulose cultures of single cells isolated from a population of CD34+ cells by fluorescence-activated cell-sorting (FACS) clone-sorting system. CD34+ cells were detected in bone marrow (BM) and peripheral blood (PB) cells at incidences of 1.0% and 0.01% of total mononuclear cells, respectively. Single cell cultures revealed that approximately 37% of BM CD34+ cells formed colonies in the presence of phytohemagglutinin-leukocyte conditioned medium and erythropoietin. Erythroid bursts-, granulocyte-macrophage (GM) colony-, and pure macrophage (Mac) colony-forming cells were 10% each in CD34+ cells. Approximately 15% of PB CD34+ cells formed colonies in which erythroid bursts were predominant. CD34+ cells were heterogeneous and fractionated by several antibodies in FACS multicolor analysis. In these fractionated cells, CD34+, CD33+ cells formed GM and Mac colonies 7 to 10 times as often as CD34+, CD33- cells. Most of the erythroid bursts and colonies were observed in the fraction of CD34+, CD13- cells or CD34+, CD33- cells. The expression of HLA-DR on CD34+ cells was not related to the incidence, size, or type of colonies. There was no difference in the phenotypical heterogeneity of CD34+ cells between BM and PB. About 10% of CD34+ cells were able to form G colonies in response to granulocyte colony-stimulating factor (G-CSF) and to form Mac colonies in GM-CSF or interleukin-3 (IL-3). Progenitors capable of generating colonies by stimulation of G-CSF were more enriched in CD34+, CD33+ fraction than in CD34+, CD33- fraction. Thus, single cell cultures using the FACS clone-sorting system provide an accurate estimation of hematopoietic progenitors and an assay system for direct action of colony-stimulating factors.

Original languageEnglish
Pages (from-to)1941-1946
Number of pages6
JournalBlood
Volume75
Issue number10
Publication statusPublished - 1990 May 15

Fingerprint

Macrophages
Sorting
Clone Cells
Bone
Blood
Fluorescence
Granulocyte Colony-Stimulating Factor
Cell culture
Granulocytes
Colony-Stimulating Factors
Methylcellulose
Flow Cytometry
Interleukin-3
Phytohemagglutinins
HLA-DR Antigens
Cell Culture Techniques
Conditioned Culture Medium
Erythropoietin
Bone Marrow Cells
Assays

ASJC Scopus subject areas

  • Hematology

Cite this

Ema, H., Suda, T., Miura, Y., & Nakauchi, H. (1990). Colony formation of clone-sorted human hematopoietic progenitors. Blood, 75(10), 1941-1946.

Colony formation of clone-sorted human hematopoietic progenitors. / Ema, Hideo; Suda, Toshio; Miura, Yasusada; Nakauchi, Hiromitsu.

In: Blood, Vol. 75, No. 10, 15.05.1990, p. 1941-1946.

Research output: Contribution to journalArticle

Ema, H, Suda, T, Miura, Y & Nakauchi, H 1990, 'Colony formation of clone-sorted human hematopoietic progenitors', Blood, vol. 75, no. 10, pp. 1941-1946.
Ema H, Suda T, Miura Y, Nakauchi H. Colony formation of clone-sorted human hematopoietic progenitors. Blood. 1990 May 15;75(10):1941-1946.
Ema, Hideo ; Suda, Toshio ; Miura, Yasusada ; Nakauchi, Hiromitsu. / Colony formation of clone-sorted human hematopoietic progenitors. In: Blood. 1990 ; Vol. 75, No. 10. pp. 1941-1946.
@article{f6f61e34a2774c7d9a73a1c92022ab59,
title = "Colony formation of clone-sorted human hematopoietic progenitors",
abstract = "To characterize human hematopoietic progenitors, we performed methylcellulose cultures of single cells isolated from a population of CD34+ cells by fluorescence-activated cell-sorting (FACS) clone-sorting system. CD34+ cells were detected in bone marrow (BM) and peripheral blood (PB) cells at incidences of 1.0{\%} and 0.01{\%} of total mononuclear cells, respectively. Single cell cultures revealed that approximately 37{\%} of BM CD34+ cells formed colonies in the presence of phytohemagglutinin-leukocyte conditioned medium and erythropoietin. Erythroid bursts-, granulocyte-macrophage (GM) colony-, and pure macrophage (Mac) colony-forming cells were 10{\%} each in CD34+ cells. Approximately 15{\%} of PB CD34+ cells formed colonies in which erythroid bursts were predominant. CD34+ cells were heterogeneous and fractionated by several antibodies in FACS multicolor analysis. In these fractionated cells, CD34+, CD33+ cells formed GM and Mac colonies 7 to 10 times as often as CD34+, CD33- cells. Most of the erythroid bursts and colonies were observed in the fraction of CD34+, CD13- cells or CD34+, CD33- cells. The expression of HLA-DR on CD34+ cells was not related to the incidence, size, or type of colonies. There was no difference in the phenotypical heterogeneity of CD34+ cells between BM and PB. About 10{\%} of CD34+ cells were able to form G colonies in response to granulocyte colony-stimulating factor (G-CSF) and to form Mac colonies in GM-CSF or interleukin-3 (IL-3). Progenitors capable of generating colonies by stimulation of G-CSF were more enriched in CD34+, CD33+ fraction than in CD34+, CD33- fraction. Thus, single cell cultures using the FACS clone-sorting system provide an accurate estimation of hematopoietic progenitors and an assay system for direct action of colony-stimulating factors.",
author = "Hideo Ema and Toshio Suda and Yasusada Miura and Hiromitsu Nakauchi",
year = "1990",
month = "5",
day = "15",
language = "English",
volume = "75",
pages = "1941--1946",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "10",

}

TY - JOUR

T1 - Colony formation of clone-sorted human hematopoietic progenitors

AU - Ema, Hideo

AU - Suda, Toshio

AU - Miura, Yasusada

AU - Nakauchi, Hiromitsu

PY - 1990/5/15

Y1 - 1990/5/15

N2 - To characterize human hematopoietic progenitors, we performed methylcellulose cultures of single cells isolated from a population of CD34+ cells by fluorescence-activated cell-sorting (FACS) clone-sorting system. CD34+ cells were detected in bone marrow (BM) and peripheral blood (PB) cells at incidences of 1.0% and 0.01% of total mononuclear cells, respectively. Single cell cultures revealed that approximately 37% of BM CD34+ cells formed colonies in the presence of phytohemagglutinin-leukocyte conditioned medium and erythropoietin. Erythroid bursts-, granulocyte-macrophage (GM) colony-, and pure macrophage (Mac) colony-forming cells were 10% each in CD34+ cells. Approximately 15% of PB CD34+ cells formed colonies in which erythroid bursts were predominant. CD34+ cells were heterogeneous and fractionated by several antibodies in FACS multicolor analysis. In these fractionated cells, CD34+, CD33+ cells formed GM and Mac colonies 7 to 10 times as often as CD34+, CD33- cells. Most of the erythroid bursts and colonies were observed in the fraction of CD34+, CD13- cells or CD34+, CD33- cells. The expression of HLA-DR on CD34+ cells was not related to the incidence, size, or type of colonies. There was no difference in the phenotypical heterogeneity of CD34+ cells between BM and PB. About 10% of CD34+ cells were able to form G colonies in response to granulocyte colony-stimulating factor (G-CSF) and to form Mac colonies in GM-CSF or interleukin-3 (IL-3). Progenitors capable of generating colonies by stimulation of G-CSF were more enriched in CD34+, CD33+ fraction than in CD34+, CD33- fraction. Thus, single cell cultures using the FACS clone-sorting system provide an accurate estimation of hematopoietic progenitors and an assay system for direct action of colony-stimulating factors.

AB - To characterize human hematopoietic progenitors, we performed methylcellulose cultures of single cells isolated from a population of CD34+ cells by fluorescence-activated cell-sorting (FACS) clone-sorting system. CD34+ cells were detected in bone marrow (BM) and peripheral blood (PB) cells at incidences of 1.0% and 0.01% of total mononuclear cells, respectively. Single cell cultures revealed that approximately 37% of BM CD34+ cells formed colonies in the presence of phytohemagglutinin-leukocyte conditioned medium and erythropoietin. Erythroid bursts-, granulocyte-macrophage (GM) colony-, and pure macrophage (Mac) colony-forming cells were 10% each in CD34+ cells. Approximately 15% of PB CD34+ cells formed colonies in which erythroid bursts were predominant. CD34+ cells were heterogeneous and fractionated by several antibodies in FACS multicolor analysis. In these fractionated cells, CD34+, CD33+ cells formed GM and Mac colonies 7 to 10 times as often as CD34+, CD33- cells. Most of the erythroid bursts and colonies were observed in the fraction of CD34+, CD13- cells or CD34+, CD33- cells. The expression of HLA-DR on CD34+ cells was not related to the incidence, size, or type of colonies. There was no difference in the phenotypical heterogeneity of CD34+ cells between BM and PB. About 10% of CD34+ cells were able to form G colonies in response to granulocyte colony-stimulating factor (G-CSF) and to form Mac colonies in GM-CSF or interleukin-3 (IL-3). Progenitors capable of generating colonies by stimulation of G-CSF were more enriched in CD34+, CD33+ fraction than in CD34+, CD33- fraction. Thus, single cell cultures using the FACS clone-sorting system provide an accurate estimation of hematopoietic progenitors and an assay system for direct action of colony-stimulating factors.

UR - http://www.scopus.com/inward/record.url?scp=0025311181&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025311181&partnerID=8YFLogxK

M3 - Article

C2 - 1692488

AN - SCOPUS:0025311181

VL - 75

SP - 1941

EP - 1946

JO - Blood

JF - Blood

SN - 0006-4971

IS - 10

ER -