Tracy Sarah A, Ahmed Azra, Tigges John C, Ericsson Maria, Pal Anoop K, Zurakowski David, Fauza Dario O
Department of Surgery, Boston Children's Hospital, Boston and Cambridge, MA.
Beth Israel Deaconess Medical Center, Boston and Cambridge, MA.
J Pediatr Surg. 2019 Jan;54(1):86-90. doi: 10.1016/j.jpedsurg.2018.10.020. Epub 2018 Oct 5.
BACKGROUND/PURPOSE: Exosomes may constitute a more practical alternative to live cells in select stem cell-based therapies. We sought to compare exosomes from two mesenchymal stem cell (MSC) sources relevant to perinatal and pediatric diseases.
Exosomes were isolated by reagent-enhanced centrifugation from cell culture media of banked human bone marrow (bm) and amniotic fluid (af) MSCs after serum starvation. Characterization was by flow exometry for tetraspanin markers CD9, CD63, and CD81, transmission electron microscopy for size and morphology, and tunable resistive pulse sensing for size distribution and concentration. Statistical comparisons of count data were made by Poisson regression modeling and Student's T-test.
Exosomes of appropriate size and morphology were isolated with comparable expressions of CD9 (96% vs. 94%), CD63 (88% vs. 66%), and CD81 (71% vs. 63%) for bmMSC and afMSC, respectively. Total exosome yield (particles/mL) adjusted for number of cells was higher from afMSCs than bmMSCs by an estimated 25% (P < 0.001).
While bone marrow and amniotic fluid mesenchymal stem cells are comparable sources of exosomes in size distribution, morphology, and expression of typical surface markers, yield may be higher from amniotic fluid cells. The amniotic fluid appears to be a preferable source of exosomes for clinical applications.
N/A (bench laboratory study).
背景/目的:在某些基于干细胞的治疗中,外泌体可能是比活细胞更实用的替代物。我们试图比较来自与围产期和儿科疾病相关的两种间充质干细胞(MSC)来源的外泌体。
在血清饥饿后,通过试剂增强离心从储存的人骨髓(bm)和羊水(af)间充质干细胞的细胞培养基中分离外泌体。通过流式细胞术检测四跨膜蛋白标志物CD9、CD63和CD81进行表征,通过透射电子显微镜观察大小和形态,通过可调电阻脉冲传感检测大小分布和浓度。计数数据的统计比较采用泊松回归模型和学生t检验。
分离出大小和形态合适的外泌体,bmMSC和afMSC的CD9(分别为96%对94%)、CD63(88%对66%)和CD81(71%对63%)表达相当。根据细胞数量调整后的外泌体总产量(颗粒/毫升),afMSC比bmMSC高约25%(P<0.001)。
虽然骨髓和羊水间充质干细胞在外泌体的大小分布、形态和典型表面标志物表达方面是可比的来源,但羊水细胞产生的外泌体产量可能更高。羊水似乎是临床应用中外泌体的更优来源。
无(实验室基础研究)。