Liu Yang, Liu Tianqing, Ma Xuehu, Fan Xiubo, Bao Chunyu, Cui Zhanfeng
Dalian R & D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian, China.
J Microencapsul. 2009 Mar;26(2):130-42. doi: 10.1080/02652040802193014.
The expansion of umbilical cord blood mononuclear cells (UCB MNCs) was investigated in a novel co-culture system by means of encapsulation of rabbit bone marrow (BM) mesenchymal stem cells (MSCs) in alginate beads (Alg beads). Three kinds of media were applied and the experiments lasted for 7 days. The total nucleated cell density was measured every 24 h. Flow cytometric assay for CD34(+) cells and methylcellulose colony assays were carried out at 0, 72 and 168 h. It was found that the encapsulated MSCs illustrated remarkable effects on UCB MNCs expansion regardless of whether serum is present in culture media or not. At the end of 168 h co-culture, the total nucleated cell number was multiplied by 15 +/- 2.9 times, and CD34(+) cells 5.3 +/- 0.3 times and colony-forming units in culture (CFU-Cs) 5.6 +/- 1.2 times in the serum-free media supplemented with conventional dose of cytokines, which was very similar to the results in the containing 20% serum media. While in the control, i.e. MNC expansion without encapsulated MSCs, however, total nucleated cells density changed mildly, CD34(+) cells and CFU-Cs showed little effective expansion. It is demonstrated that the encapsulated stromal cells can support the expansion of UCB MNCs effectively under the experimental condition.
通过将兔骨髓(BM)间充质干细胞(MSCs)包裹在藻酸盐珠(Alg珠)中,在一种新型共培养系统中研究了脐带血单个核细胞(UCB MNCs)的扩增情况。应用了三种培养基,实验持续7天。每24小时测量一次有核细胞总数密度。在0、72和168小时进行CD34(+)细胞的流式细胞术检测和甲基纤维素集落检测。结果发现,无论培养基中是否存在血清,包裹的MSCs对UCB MNCs的扩增均显示出显著效果。在共培养168小时结束时,在补充常规剂量细胞因子的无血清培养基中,有核细胞总数增加了15±2.9倍,CD34(+)细胞增加了5.3±0.3倍,培养中的集落形成单位(CFU-Cs)增加了5.6±1.2倍,这与含20%血清培养基中的结果非常相似。然而,在对照实验中,即没有包裹MSCs的MNCs扩增实验中,有核细胞总数密度变化不大,CD34(+)细胞和CFU-Cs几乎没有有效扩增。结果表明,在实验条件下,包裹的基质细胞可以有效地支持UCB MNCs的扩增。