聚己内酯纳米纤维支架体外扩增脐血CD34+造血干细胞的潜力

Mousavi Seyed Mohammad Hadi, A. Saeid, S. Masoud, M. Javad
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引用次数: 4

摘要

背景:高效、实用的脐带血造血干细胞体外扩增作为造血干细胞移植的替代来源,对于理解造血干细胞移植在多种血液学和非血液学疾病的治疗或支持治疗中的潜力至关重要。这项研究的目的是脐带血造血干细胞体外扩增在一个新的三维聚己内酯纳米纤维支架涂有胶原蛋白。方法:将脐带血CD34+细胞分别在二维和三维培养体系中培养10 d后,采用qRT-PCR、流式细胞术和克隆原性等方法进行评价。结果:胶原包被的三维聚己内酯纳米支架提供了更高的总有核细胞(50倍比38倍)和CD34+细胞(20倍比2.6倍)(p < 0.05),与二维细胞培养和扩增前相比,归巢和自我更新基因的表达更高,VLA-4、hTERT和BMI-1基因的表达有统计学意义(p = 0.0001)。骨髓标志物在三维支架中的表达显著高于二维培养体系(p < 0.05)。二维培养基中菌落总数低于三维培养基(p < 0.05)。结论:本研究证明了支架的三维性与作为细胞外基质蛋白的胶原之间的协同作用,以及胶原包被的三维聚己内酯纳米纤维支架体外扩增造血干细胞的潜力。
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Potential of Polycaprolactone Nanofiber Scaffold for Ex Vivo Expansion of Cord Blood-Derived CD34+ Hematopoietic Stem Cells
Background: An efficient and practical ex vivo expansion of cord blood hematopoietic stem cells as an alternative source of HSC transplantation is crucial in understanding the potential of HSC transplantation in treating or supportive therapy in a variety of hematologic and non-hematologic disorders. The aim of this study was an ex vivo expansion of cord blood hematopoietic stem cells in a novel threedimensional polycaprolactone nanofiber scaffold coated with collagen. Methods: After 10-day cultured of cord blood CD34+ cells in 2-Dimensional and 3-Dimensional culture system, the evaluation was performed by qRT-PCR, flow cytometry and clonogenicity. Results: 3-Dimensional Polycaprolactone nano-scaffold coated with collagen provided higher total nucleated cells (50-fold vs. 38-fold) and CD34+ cells (20-fold vs. 2.6-fold) (p < 0.05) and compared to 2-Dimensional cell culture and before expansion had higher expression of homing and self-renewal genes and for VLA-4, hTERT and BMI-1 genes were statically significant (p = 0.0001). The expression of myeloid markers in 3-Dimensional scaffold was significantly higher than the 2-Dimensional culture system (p < 0.05). The total colony in 2-Dimensional culture was lower than 3-Dimensional culture medium (p < 0.05). Conclusion: This study demonstrated the synergistic effect between the three-dimensionality of the scaffold and collagen as an extracellular matrix protein, and the potential of this 3-Dimensional Polycaprolactone nanofiber scaffold coated with collagen for ex vivo expansion of HSCs.
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