Erythroid differentiation and maturation from peripheral CD34+ cells in liquid culture:: Cellular and molecular characterization

被引:37
|
作者
Ronzoni, Luisa [1 ]
Bonara, Paola [1 ]
Rusconi, Daniela [1 ]
Frugoni, Cecilia [1 ]
Libani, Ilaria [1 ]
Cappellini, Maria Domenica [1 ]
机构
[1] Univ Milan, Dept Internal Med, IRCCS, Fdn Policlin Mangiagalli, I-20122 Milan, Italy
关键词
erythroid differentiation; erythropoietin; CD34(+) peripheral blood cells; flow cytometric analysis; erythroid gene expression;
D O I
10.1016/j.bcmd.2007.07.006
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
In vitro models of human erythropoiesis are useful in studying the mechanisms of erythroid differentiation from BFU-E to mature erythrocytes both in normal and pathological conditions. Most of the available in vitro liquid cultures are from cell lines or are limited by the production of few erythroid cells mixed with myeloid cells. Here we describe an erythroid liquid culture system starting from CD34(+)-enriched cells obtained from peripheral blood. CD34(+) cells were cultured for 21 days in different conditions. Precisely stem cell factor (SCF, 20 ng/mL) and IL-3 (10 ng/mL) were added at starting point plus Epo (3 U/mL) at day 0 or 7 of culture with or without cyclosporine A (Cy; 1 mg/mL). In all the conditions, the highest recovery was obtained at day 14 of culture. Epo and Cy added at day 0 produced the highest cell expansion (170-fold mean amplification of the initial cell input by day 14) and recovery of erythroid cell. Sixty seven percent of the cells were GP(+) at day 7 and 97% by day 14 respectively. Most of the cells were proerythroblasts at day 7 and mature erythroblasts at day 14 (> 90% were benzidine(pos)). The presence of Cy favoured erythroid differentiation and maturation and reduced the percentage of non-erythroid CD45(+) cells (2% with Cy versus 5% without Cy). Cells cultured with Epo and Cy reproduced erythropoiesis also at the molecular level. The results suggest that in 14 days different steps of human erythropoiesis from peripheral CD34+ cells could be reproduced, with high recovery of highly purified erythroid cells. The high number and purity of erythroid cells produced from a small amount of peripheral blood make this method useful for studying either normal or pathological erythropoiesis. (c) 2007 Elsevier Inc. All rights reserved.
引用
收藏
页码:148 / 155
页数:8
相关论文
共 50 条
  • [31] Characterization of Splenic CD34+ Cells From Patients with Primary Myelofibrosis
    Wang, Xiaoli
    Prakash, Sonam
    Lu, Min
    Li, Yan
    Orazi, Attilio
    Hoffman, Ronald
    BLOOD, 2011, 118 (21) : 1211 - 1212
  • [32] Characterization of CD34+ cells isolated from human fetal lung
    Acarregui, MJ
    England, KM
    Richman, JT
    Littig, JL
    AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 2003, 284 (02) : L395 - L401
  • [33] Differentiation of cancer induced CD34+ cells into dendritic cells.
    Lathers, DMR
    Lubbers, E
    Beal, NM
    Wright, MA
    Young, MRI
    FASEB JOURNAL, 1999, 13 (04): : A301 - A301
  • [34] A serum-free culture model for studying the differentiation of human dendritic cells from adult CD34+ progenitor cells
    Luft, T
    Pang, KC
    Thomas, E
    Bradley, CJ
    Savoia, H
    Trapani, J
    Cebon, J
    EXPERIMENTAL HEMATOLOGY, 1998, 26 (06) : 489 - 500
  • [35] Culture of CD34+ cells from placental/umbilical cord blood.
    Pranke, P
    Hendrikx, J
    Debnath, G
    Alespeiti, G
    Rubinstein, P
    Visser, JWM
    BLOOD, 2001, 98 (11) : 658A - 658A
  • [36] Docetaxel effectively mobilizes peripheral blood CD34+ cells
    Prince, HM
    Toner, GC
    Seymour, JF
    Blakey, D
    Gates, P
    Eerhard, S
    Chapple, P
    Wall, D
    Quinn, M
    Juneja, S
    Wolf, M
    Januszewicz, EH
    Richardson, G
    Scarlett, J
    Briggs, P
    Brettell, M
    Rischin, D
    BONE MARROW TRANSPLANTATION, 2000, 26 (05) : 483 - 487
  • [37] Docetaxel effectively mobilizes peripheral blood CD34+ cells
    HM Prince
    GC Toner
    JF Seymour
    D Blakey
    P Gates
    S Eerhard
    P Chapple
    D Wall
    M Quinn
    S Juneja
    M Wolf
    EH Januszewicz
    G Richardson
    J Scarlett
    P Briggs
    M Brettell
    D Rischin
    Bone Marrow Transplantation, 2000, 26 : 483 - 487
  • [38] Osteoprotegerin Induces CD34+ Differentiation in Endothelial Progenitor Cells
    Boisson-Vidal, Catherine
    Benslimane-Ahmim, Zahia
    Lokajczyk, Anna
    Heymann, Dominique
    Smadja, David M.
    FRONTIERS IN MEDICINE, 2018, 5
  • [39] Identification and functional characterization of ion channels in CD34+ hematopoietic stem cells from human peripheral blood
    Park, Kyoung Sun
    Pang, Bo
    Park, Su Jung
    Lee, Yun-Gyoo
    Bae, Ji-Yeon
    Park, Seonyang
    Kim, Inho
    Kim, Sung Joon
    MOLECULES AND CELLS, 2011, 32 (02) : 181 - 188
  • [40] Differentiation of CD34+ cord blood cells in glycosaminoglycan microcapsules
    Buruzula, GK
    Matthew, HWT
    Eliason, JF
    PROCEEDINGS OF THE IEEE 30TH ANNUAL NORTHEAST BIOENGINEERING CONFERENCE, 2004, : 114 - 115