Expansion of HPCs from cord blood in a novel 3D matrix
Expansion of HPCs from cord blood in a novel 3D matrix
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DOI：
10.1080/14653240310003585
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发表时间：
2003-12-01
期刊：
影响因子：
4.5
通讯作者：
Rosenzweig, M
中科院分区：
文献类型：
--
作者：
Ehring, B;Biber, K;Rosenzweig, M
背景 用于扩展多能 HPC 的最佳系统将理想地消除细胞因子和动物源血清的使用。我们之前已经证明，3D 钽涂层多孔生物材料 (Cytomatrix) 在没有细胞因子的情况下支持人类骨髓 HPC 的维持和扩增。方法在没有外源细胞因子的情况下，在 Cytomatrix 中培养脐带血 (UCB) 衍生的 HPC。使用FACS测定表型。评估菌落形成单位（CFU）活性。通过将扩增的细胞移植到非肥胖糖尿病 (NOD)/SCID 小鼠中来评估植入能力。结果我们描述了使用 Cytomatrix 系统从 UCB 中扩增 HPC。当 UCB 衍生的 CD34(+) 细胞在 Cytomatrix 系统中培养 2 周时，我们观察到有核细胞数量（3 倍）和 CFU（2.6 倍）增加。 CD45(+)和CD34(+)细胞的数量均增加了三倍。趋势表明CD34(+) C38(-)细胞的频率增加，CD34(+) C33(+)细胞和CD34(+) C61(+)细胞的频率增加。未观察到 T 或 B 淋巴细胞的扩增。当将来自 Cytomatrix 的扩增 UCB 细胞注射到亚致死照射的 NOD/SCID 小鼠中时，移植后 6 周，在小鼠外周血和 BM 中检测到人类细胞。讨论 这种在无血清、无细胞因子的环境中扩增 UCB 细胞的独特方法可能会提供具有多谱系移植能力的 HPC 扩增，可用于临床。
Background An optimal system for the expansion of pluripotent HPCs would ideally eliminate the use of cytokines and animal-derived serum. We have shown previously that a 3D, tantalum-coated porous biomaterial ( Cytomatrix) supports the maintenance and expansion of human BM HPCs in the absence of cytokines.Methods Umbilical cord blood (UCB) derived HPC were cultured in the Cytomatrix in the absence of exogenous cytokines. Phenotype was determined using FACS. Colony-forming units (CFU) activity was evaluated. Engraftment capacity was evaluated by transplanting the expanded cells into non-obese diabetic (NOD)/SCID mice.Results We describe the expansion of HPCs from UCB using the Cytomatrix system. When UCB-derived CD34(+) cells were cultured in the Cytomatrix system for 2 weeks we observed an increase in the number of nucleated cells (3-fold) and CFU (2.6-fold). The number of CD45(+) and CD34(+) cells both increased three-fold. Trends demonstrated an increase in the frequency of CD34(+) C38(-) cells, and an increase in both CD34(+) C33(+) cells and CD34(+) C61(+) cells. No expansion of T or B lymphocytes was observed. When expanded UCB cells from the Cytomatrix were injected into sub-lethally irradiated NOD/SCID mice, human cells were detected in the murine peripheral blood and BM 6 weeks post-transplantation.Discussion This unique approach to the expansion of UCB cells in a serum-free, cytokine-free environment may provide expansion of HPCs with multi-lineage engraftment capability that could be used clinically.