Kögler Gesine, Radke Teja Falk, Lefort Aurélie, Sensken Sandra, Fischer Johannes, Sorg Rüdiger V, Wernet Peter
Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Düsseldorf, Germany.
Exp Hematol. 2005 May;33(5):573-83. doi: 10.1016/j.exphem.2005.01.012.
Cytokine production and hematopoiesis-supporting stromal activity of cord blood (CB)-derived unrestricted somatic stem cells (USSC) in comparison to bone marrow mesenchymal stem cells (BMMSC) and hematopoietic progenitor expansion solely driven by recombinant cytokines were assessed.
USSC generation was initiated from fresh and cryopreserved CB. Cytokine production by USSC and BMMSC was determined qualitatively by cytokine mRNA expression array analyses or quantitatively by Multiplex or ELISA analyses. To evaluate hematopoiesis-supporting activity, CB CD34+ cells were expanded in cocultures with USSC and BMMSC or in the presence of Flt3-L, SCF, and TPO. Expansion of CD34+ cells, total cells, colony-forming cells (CFC), and LTC-IC were determined after 1, 2, 3, and 4 weeks of culture.
USSC constitutively produced SCF, LIF, TGF-1beta, M-CSF, GM-CSF, VEGF, IL-1beta, IL-6, IL-8, IL-11, IL-12, IL-15, SDF-1alpha, and HGF. When USSC were stimulated with IL-1beta, G-CSF was released. Production of SCF and LIF were significantly higher in USSC compared to BMMSC. At 1, 2, 3, and 4 weeks, cocultivation of CD34+ cells on the USSC layer resulted in a 14.6-fold +/- 1.1-fold, 110.1-fold +/- 17.9-fold, 151.8-fold +/- 39.7-fold, and 183.6-fold +/- 40.4-fold amplification of total cells and in a 30.6-fold +/- 4.4-fold, 101.4-fold +/- 27.5-fold, 64.7-fold +/- 15.8-fold, and 29.4-fold +/- 3.1-fold amplification of CFC, respectively. LTC-IC expansion at 1 and 2 weeks was, with 2.0-fold +/- 0.1-fold and 2.5-fold +/- 0.3-fold, significantly higher for USSC than BMMSC (1.1-fold +/- 0.03-fold and 1.1-fold +/- 0.1-fold), but declined after day 21. Transwell cocultures of USSC did not significantly alter total cell or CFC expansion.
USSC produce functionally significant amounts of hematopoiesis-supporting cytokines and are superior to BMMSC in expansion of CD34+ cells from CB. USSC is therefore a suitable candidate for stroma-driven ex vivo expansion of hematopoietic CB cells for short-term reconstitution.
评估与骨髓间充质干细胞（BMMSC）相比，脐血（CB）来源的无限制体细胞干细胞（USSC）的细胞因子产生情况以及造血支持基质活性，同时评估仅由重组细胞因子驱动的造血祖细胞扩增情况。
从新鲜和冷冻保存的CB中获取USSC。通过细胞因子mRNA表达阵列分析定性或通过多重分析或酶联免疫吸附分析（ELISA）定量测定USSC和BMMSC产生的细胞因子。为评估造血支持活性，将CB CD34+细胞与USSC和BMMSC共培养或在存在Flt3-L、干细胞因子（SCF）和血小板生成素（TPO）的情况下进行扩增。在培养1、2、3和4周后测定CD34+细胞、总细胞、集落形成细胞（CFC）和长期培养起始细胞（LTC-IC）的扩增情况。
USSC持续产生SCF、白血病抑制因子（LIF）、转化生长因子-1β（TGF-1β）、巨噬细胞集落刺激因子（M-CSF）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、血管内皮生长因子（VEGF）、白细胞介素-1β（IL-1β）、IL-6、IL-8、IL-11、IL-12、IL-15、基质细胞衍生因子-1α（SDF-1α）和肝细胞生长因子（HGF）。当用IL-1β刺激USSC时，会释放粒细胞集落刺激因子（G-CSF）。与BMMSC相比，USSC中SCF和LIF的产生显著更高。在第1、2、3和4周时，将CD34+细胞在USSC层上共培养导致总细胞扩增了14.6倍±1.1倍、110.1倍±17.9倍、151.8倍±39.7倍和183.6倍±40.4倍，CFC扩增了30.6倍±4.4倍、101.4倍±27.5倍、64.7倍±15.8倍和29.4倍±3.1倍。在第1周和第周时，USSC的LTC-IC扩增分别为2.0倍±0.1倍和2.5倍±0.3倍，显著高于BMMSC（1.1倍±0.03倍和1.1倍±0.1倍），但在第21天后下降。USSC的Transwell共培养未显著改变总细胞或CFC扩增。
USSC产生功能上显著量的造血支持细胞因子，并且在扩增CB来源的CD34+细胞方面优于BMMSC。因此，USSC是用于基质驱动的造血CB细胞体外短期重建扩增的合适候选者。
