包埋的脐血cd34细胞与仿生壁龛的动态共培养化学工程专业论文.docxVIP

包埋的脐血CD34+细胞与仿生壁龛的动态共培养来研究动态环境和仿生壁龛对CD34+细胞的作用。通过计算包埋收获的CD34+细胞的增 包埋的脐血CD34+细胞与仿生壁龛的动态共培养 来研究动态环境和仿生壁龛对CD34+细胞的作用。通过计算包埋收获的CD34+细胞的增 殖倍数、活率得出，第10天时，动态共培养条件下，CD34+细胞扩增24．2±2．3倍，并 且可以维持84．1％的活率，而两个静态组中，特别是无仿生壁龛支持的情况下，细胞扩 增缓慢，且只有约50％的活率。通过流式测定CD34表达率、集落形成单元计数，发现 在三组实验中，CD34的表达率都会下降，但动态实验组下降最慢，在10天时，CD34 仍可维持80．3％的表达率；同时，动态实验组细胞的集落形成单元有明显的扩增，在第 14天可以扩增11．7±2．5倍，而在静态组的扩增倍数则较低。以上结果，可以充分证明 动态环境和仿生壁龛可以共同促进脐血CD34+细胞的增殖，并维持细胞的活性和干性。 本文构建了一种新的扩增体系，即动态环境+仿生壁龛+包埋基质体系，来扩增脐血 CD34+细胞。该体系不仅强化了营养物质或因子的传递，为CD34+细胞提供及时的营养， 同时也通过仿生壁龛为CD34+细胞提供了类体内的微环境，使得CD34+细胞在实现扩增 的同时，也可以维持一定的干性。基于基质的包埋隔离，扩增收获的CD34+细胞有望作 为有效的移植体应用于临床。 关键词：脐血CD34+细胞；仿生成骨细胞壁龛；CAG胶珠；动态环境；隔离共培养 万方数据 大连理工大学硕士学位论文Dynamic 大连理工大学硕士学位论文 Dynamic coculture of embedded UCB-CD34+cell and biomimetic niche Abstract Umbilical cord blood(UCB)as valuable biological resource，has been applied to treat hematological malignancy，solid tumor，genetic disease and SO on，with its transplatation advantages and abundant HSPCs．However,one UCB unit wit}l lower HSPCs，limits its application in clinical．So it was an important strategy to expand UCB—HSPCs by engineered procedures in vitro．Currently,two main problems exist in traditional expansion methods：1) HSCs are inclined to differentiate earlier to lose stemness；2)Allogenic stromal cells and exogenous factors mixed with expanded HSCs to result in graft failure．W11ile．it is found that in vivo bone marrow microenvironment allows HSCs to self-renew for a life span． Therefore，we first fabricate a biomimetic osteoblastic niche to provide an indirect support for HSPCs，then prepare an excellent gel—bead for embedding stromal cells to isolate allogenic stromal cells，finally expand embedded UCB—CD34+cell by biomimetic niche under dynamic condition． Fabrication of biomimetic osteoblastic niche：Firstly，HAp—CS／Gel scaffolds with varied ratio are prepared by freeze drying and chemical crosslink and it is found that HAp doping doesn’t destroy the morphology of scaffolds，which feature interconnected pores with siz