a Department of Hematology, Faculty of Allied Medical Sciences , Tehran University of Medical Sciences , Tehran , Iran.
b Department of Hematology, Faculty of Medical Sciences , Tarbiat Modares University , Tehran , Iran.
Artif Cells Nanomed Biotechnol. 2018 Jun;46(4):740-748. doi: 10.1080/21691401.2017.1337026. Epub 2017 Jul 7.
Cord blood (CB) haematopoietic stem cell (HSC) is an alternative source of HSC transplantation. The limited cell number greatly restricts their clinic-scale therapeutic applications. The objective of this study was an ex vivo expansion of CB HSCs in a new three-dimensional polycaprolactone nano-scaffold coated with fibronectin (FN). First, we isolated CB CD34+ cells and cultured 10 days in presence of growth factors. The evaluation was performed by qRT-PCR, flow cytometry and clonogenicity. 3D PCL nano-scaffold coated with FN produced significantly higher total nucleated cells and CD34+ cells (p < .05) and also had significantly higher homing and self-renewality genes than 2D cell culture and before expansion (p < .05). The expression of CXCR-4, VLA-4, VLA-5 and LFA-1, and also HOXB-4, HOXA-9, BMI-1 and hTERT genes was higher in 3D than 2D. The CD13, CD14, CD33, CD34 and CD45 markers were significantly higher and CD2, CD3 and CD19 markers were significantly lower in 3D scaffold than 2D cell culture (p < .05). The type and number of colonies in 2D culture were lower than 3D culture medium (p > .05). 3D PCL nano-scaffold coated with FN could better keep specifications homing and self renewality of CB HSCs after expansion.
脐带血(CB)造血干细胞(HSC)是 HSC 移植的替代来源。有限的细胞数量极大地限制了它们在临床规模上的治疗应用。本研究的目的是在涂有纤维连接蛋白(FN)的新型三维聚己内酯纳米支架上体外扩增 CB HSCs。首先,我们分离 CB CD34+细胞,并在存在生长因子的情况下培养 10 天。通过 qRT-PCR、流式细胞术和克隆形成进行评估。涂有 FN 的 3D PCL 纳米支架可产生显著更高的总核细胞和 CD34+细胞(p<.05),并且与 2D 细胞培养和扩增前相比,具有更高的归巢和自我更新基因(p<.05)。CXCR-4、VLA-4、VLA-5 和 LFA-1 以及 HOXB-4、HOXA-9、BMI-1 和 hTERT 基因的表达在 3D 中高于 2D。3D 支架中的 CD13、CD14、CD33、CD34 和 CD45 标志物明显高于 2D 细胞培养,CD2、CD3 和 CD19 标志物明显低于 2D 细胞培养(p<.05)。2D 培养中的集落类型和数量低于 3D 培养基(p>.05)。涂有 FN 的 3D PCL 纳米支架可更好地保持 CB HSCs 扩增后的归巢和自我更新特性。