Expansion of CD34+ Cells on Telomerized Human Stromal Cells without Losing Erythroid-Differentiation Potential in a Serum-Free Condition
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Erythropoiesis progresses from stem cell expansion on stromal cells through the formation of an erythroblastic island. Our aim was to assess the feasibility of using human stromal cells for erythroid production and differentiation. When cord blood CD34+ cells were cocultured with telomerized human stromal cells (hTERT-stromal cells) for 2 weeks, the CD34+ cells and burst-forming units-erythroid (BFU-E) significantly expanded, and a few hematopoietic cells transmigrated below the stromal layer. When nonadherent hematopoietic progenitor cells that had expanded above the hTERT-stromal cells (group B) were collected and subjected to our erythroid-differentiation protocol, they differentiated into erythroblasts with a slight hemoglobin synthesis. When the few hematopoietic cells that had transmigrated below the stromal layer were expanded for an additional 2 to 6 weeks, they exhibited a cobblestone-like appearance, and a large amount of BFU-E clambered weekly from the underside of the stromal layer to above the stromal layer (group C).When the hematopoietic progenitor cells in group C were subjected to the erythroid-differentiation protocol, large numbers of mature erythroblasts (more than 300,000 times the initial CD34+ cell number) were produced. Our hTERT-stromal expansion protocol may contribute to the construction of a system for large-scale, long-term production of erythroid cells.
Key wordsErythropoiesis Human stromal cells Erythroblast
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- 2.Kapur R, Cooper R, Zhang L, Williams DA. Cross-talk between α4β1/α5β1 and c-Kit results in opposing effect on growth and survival of hematopoietic cells via the activation of focal adhesion kinase, mitogen-activated protein kinase, and Akt signaling pathways. Blood. 2001;97:1975–1981.PubMedCrossRefGoogle Scholar
- 19.Furusawa T, Yanai N, Hara T, Miyajima A, Obinata M. Integrinassociated protein (IAP, also termed CD47) is involved in stromasupported erythropoiesis. J Biochem (Tokyo). 1998;123:101–106.Google Scholar
- 20.Sato Y, Hong HN, Yanai N, Obinata M. Involvement of stromal membrane-associated protein (SMAP-1) in erythropoietic microenvironment. J Biochem (Tokyo). 1998;124:209–216.Google Scholar
- 30.Kobune M, Xu Y, Baum C, Kelley MR, Williams DA. Retrovirusmediated expression of the base excision repair proteins, formamidopyrimidine DNA glycosylase or human oxoguanine DNA glycosylase, protects hematopoietic cells from N,Nβ,Nα- triethylenethiophosphoramide (thioTEPA)-induced toxicity in vitro and in vivo. Cancer Res. 2001;61:5116–5125.PubMedGoogle Scholar
- 32.Dai MS, Heinrich MC, Broxmeyer HE, Lu L. Enhancing effects of co-transduction of both human erythropoietin receptor and c-kit cDNAs into hematopoietic stem/progenitor cells from cord blood on proliferation and differentiation of erythroid progenitors. Cytokines Cell Mol Ther. 2000;6:1–8.PubMedCrossRefGoogle Scholar
- 37.Papadaki HA, Kritikos HD, Valatas V, Boumpas DT, Eliopoulos GD. Anemia of chronic disease in rheumatoid arthritis is associated with increased apoptosis of bone marrow erythroid cells: improvement following anti-tumor necrosis factor-_ antibody therapy. Blood. 2002;100:474–482.PubMedCrossRefGoogle Scholar