Biopharmaceutical R&D Center, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou, 215126, China.
Biopharmagen Corp., Suzhou, 215126, China.
Stem Cell Res Ther. 2018 Jan 22;9(1):14. doi: 10.1186/s13287-018-0769-5.
Autologous transplantation of endothelial progenitor cells (EPCs) is a promising therapeutic approach in the treatment of various vascular diseases. We previously reported a two-step culture system for scalable generation of human EPCs derived from cord blood CD34 cells ex vivo. Here, we now apply this culture system to expand and differentiate human and nonhuman primate EPCs from mobilized peripheral blood (PB) CD34 cells for the therapeutic potential of autologous transplantation.
The human and nonhuman primate EPCs from mobilized PB CD34 cells were cultured according to our previously reported system. The generated adherent cells were then characterized by the morphology, surface markers, nitric oxide (NO)/endothelial NO synthase (eNOS) levels and Dil-acetylated low-density lipoprotein (Dil-Ac-LDL) uptake/fluorescein isothiocyanate (FITC)-lectin binding actives. Furthermore, the efficacy and safety studies were performed by autologous transplantation via hepatic portal vein injection in a nonhuman primate model with acute liver sinusoidal endothelial cell injury.
The mobilized PB CD34 cells from both human and nonhuman primate were efficiently expanded and differentiated. Over 2 × 10 adherent cells were generated from 20 mL mobilized primate PB (1.51 × 10 ± 3.39 × 10 CD34 cells) by 36-day culture and more than 80% of the produced cells were identified as EPCs/endothelial cells (ECs). In the autologous transplant model, the injected EPC/ECs from nonhuman primate PB were scattered in the intercellular spaces of hepatocytes at the hepatic tissues 14 days post-transplantation, indicating successful migration and reconstitution in the liver structure as the functional EPCs/ECs.
We successfully applied our previous two-step culture system for the generation of primate EPCs from mobilized PB CD34 cells, evaluated the phenotypes ex vivo, and transplanted autologous EPCs/ECs in a nonhuman primate model. Our study indicates that it may be possible for these ex-vivo high-efficient expanded EPCs to be used in clinical cell therapy.
自体移植内皮祖细胞（EPCs）是治疗各种血管疾病的一种很有前途的治疗方法。我们之前报道了一种两步培养系统，可从脐带血 CD34 细胞体外大规模生成人类 EPCs。在这里，我们现在应用该培养系统从动员后的外周血（PB）CD34 细胞中扩增和分化人类和非人类灵长类动物的 EPCs，以实现自体移植的治疗潜力。
根据我们之前报道的系统，从动员后的 PB CD34 细胞中培养人类和非人类灵长类动物的 EPCs。然后通过形态学、表面标志物、一氧化氮（NO）/内皮型一氧化氮合酶（eNOS）水平和 Dil-乙酰化低密度脂蛋白（Dil-Ac-LDL）摄取/荧光素异硫氰酸酯（FITC）-植物血凝素结合活性来对生成的贴壁细胞进行表征。此外，通过非人类灵长类动物急性肝窦内皮细胞损伤模型中的肝门静脉注射进行自体移植，对其进行疗效和安全性研究。
人类和非人类灵长类动物的动员 PB CD34 细胞均能有效扩增和分化。通过 36 天的培养，从 20 mL 动员的灵长类 PB（1.51×10±3.39×10 CD34 细胞）中可生成超过 2×10 个贴壁细胞，产生的细胞中超过 80%被鉴定为 EPC/内皮细胞（ECs）。在自体移植模型中，移植的非人类灵长类动物 PB 来源的 EPC/ECs 在肝组织中 14 天移植后散布在肝细胞的细胞间隙中，表明其作为功能性 EPC/ECs 在肝脏结构中成功迁移和重建。
我们成功地将我们之前的两步培养系统应用于从动员后的 PB CD34 细胞中生成灵长类动物的 EPCs，对其进行了体外表型评估，并在非人类灵长类动物模型中移植了自体 EPC/ECs。我们的研究表明，这些体外高效扩增的 EPCs 可能可用于临床细胞治疗。
