体外培养骨髓单个核细胞分泌促血管生长因子的实验研究

Experimental study of angiogenin factors secreted by bone marrow mononuclear cells cultured in vitro

目的分析体外培养的骨髓单个核细胞持续分泌促血管生长因子的能力。方法从大鼠胫骨及股骨采集骨髓,密度梯度离心法分离出骨髓单个核细胞进行体外培养,并连续收集4周培养上清液。酶联免疫吸附实验(ELISA)法测定培养上清液中碱性成纤维细胞生长因子(bFGF)、血管内皮细胞生长因子(VEGF)和白介素-1β(IL-1β)等因子水平。结果第1、2、3、4周骨髓单个核细胞体外培养上清液中VEGF分别为(24.40±7.99)pg/m、l(89.28±5.13)pg/m、l(115.24±10.08)pg/m、l(157.00±15.64)pg/m l;bFGF含量分别为(52.72±2.13)pg/m、l(48.10±6.41)pg/m、l(44.71±3.21)pg/m、l(25.61±2.42)pg/m l;IL-1β含量分别为:(31.28±5.44)pg/m、l(71.87±3.01)pg/m、l(55.77±11.94)pg/m、l(41.75±9.14)pg/m。l结论体外培养骨髓单个核细胞可持续分泌VEGF、bFGF、IL-1β等多种促血管生长因子。

Objective To define the secretory capability of angiogenin factors by bone marrow mononuclear cells （BM-MNC） cultured in vitro. Methods Rat BM-MNCs were isolated by centrifugation through a Histopaque density gradient and cultured for four weeks in vitro. The levels of vascular endothelial cell growth factor （VEGF）, basic fibroblast growth factor （bFGF） and interleukin-1β（IL-1β） were tested by enzyme - linked immunosorbent assay （ELISA）. Results The levels of VEGF in the conditioned medium at 1,2, 3 and 4 weeks were （24. 40 ± 7.99）, （89. 28 ± 5. 13 ）, （ 115.24 ± 10. 08） and （ 157.00 ±15.64） pg/ml, respectively. The levels of bFGF at 1, 2, 3 and 4 weeks were （52. 72±2. 13）, （48. 10±6. 41 ）, （44. 71±3. 21 ） and （25.61±2. 42） pg/ml respectively, and those of IL-1β were （31.28±5.44）, （71.87±3.01） , （55.77±11.94） and （41.75±9.14）, respectively. Conclusion The BM-MNCs can persistently secrete potent angiogenie cytokines such as VEGF, bFGF and IL-1β.