Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore, Kent Ridge, Singapore 1190741, Singapore.
J Cell Biochem. 2012 Feb;113(2):658-68. doi: 10.1002/jcb.23395.
Bone marrow mesenchymal stromal cells (BMMSCs) have been used as feeder support for the ex vivo expansion of hematopoietic stem cells (HSCs) but have the limitations of painful harvest, morbidity, and risk of infection to the patient. This prompted us to explore the use of human umbilical cord Wharton's jelly MSCs (hWJSCs) and its conditioned medium (hWJSC-CM) for ex vivo expansion of HSCs in allogeneic and autologous settings because hWJSCs can be harvested in abundance painlessly, are proliferative, hypoimmunogenic, and secrete a variety of unique proteins. In the presence of hWJSCs and hWJSC-CM, HSCs put out pseudopodia-like outgrowths and became highly motile. Time lapse imaging showed that the outgrowths helped them to migrate towards and attach to the upper surfaces of hWJSCs and undergo proliferation. After 9 days of culture in the presence of hWJSCs and hWJSC-CM, MTT, and Trypan blue assays showed significant increases in HSC numbers, and FACS analysis generated significantly greater numbers of CD34(+) cells compared to controls. hWJSC-CM produced the highest number of colonies (CFU assay) and all six classifications of colony morphology typical of hematopoiesis were observed. Proteomic analysis of hWJSC-CM showed significantly greater levels of interleukins (IL-1a, IL-6, IL-7, and IL-8), SCF, HGF, and ICAM-1 compared to controls suggesting that they may be involved in the HSC multiplication. We propose that cord blood banks freeze autologous hWJSCs and umbilical cord blood (UCB) from the same umbilical cord at the same time for the patient for future ex vivo HSC expansion and cell-based therapies.
骨髓间充质基质细胞 (BMMSCs) 已被用作体外扩增造血干细胞 (HSCs) 的饲养支持物，但存在疼痛采集、发病率和患者感染风险等限制。这促使我们探索使用人脐带华通氏胶间充质干细胞 (hWJSCs) 及其条件培养基 (hWJSC-CM) 进行异体和自体环境下的 HSCs 体外扩增，因为 hWJSCs 可以大量无痛采集，增殖能力强，免疫原性低，并分泌多种独特的蛋白质。在 hWJSCs 和 hWJSC-CM 的存在下，HSCs 伸出伪足样突起并变得高度运动。延时成像显示，这些突起帮助它们向 hWJSCs 上表面迁移并附着，并发生增殖。在 hWJSCs 和 hWJSC-CM 的存在下培养 9 天后，MTT 和台盼蓝检测显示 HSC 数量显著增加，流式细胞术分析产生的 CD34(+)细胞数量明显多于对照。hWJSC-CM 产生的集落数量最多（CFU 检测），观察到所有六种典型造血的集落形态分类。hWJSC-CM 的蛋白质组学分析显示，白细胞介素 (IL-1a、IL-6、IL-7 和 IL-8)、SCF、HGF 和 ICAM-1 的水平明显高于对照，表明它们可能参与 HSC 增殖。我们提出，脐带血库同时为患者冷冻自体 hWJSCs 和来自同一脐带的脐带血，以备将来进行体外 HSC 扩增和基于细胞的治疗。
