Joseph Matthew, Das Manjusri, Kanji Suman, Lu Jingwei, Aggarwal Reeva, Chakroborty Debanjan, Sarkar Chandrani, Yu Hongmei, Mao Hai-Quan, Basu Sujit, Pompili Vincent J, Das Hiranmoy
Stem Cell Research Laboratory, Davis Heart and Lung Research Institute, Wexner Medical Center at The Ohio State University, Columbus, OH, USA.
Department of Pathology, Arthur G. James Cancer Hospital and Richard J. Solove Research Institute, The Ohio State University Comprehensive Cancer Center, Columbus, OH, USA.
Biomaterials. 2014 Oct;35(30):8566-75. doi: 10.1016/j.biomaterials.2014.06.037. Epub 2014 Jul 4.
Despite recent advances in cardiovascular medicine, ischemic diseases remain a major cause of morbidity and mortality. Although stem cell-based therapies for the treatment of ischemic diseases show great promise, limited availability of biologically functional stem cells mired the application of stem cell-based therapies. Previously, we reported a PES-nanofiber based ex vivo stem cell expansion technology, which supports expansion of human umbilical cord blood (UCB)-derived CD133(+)/CD34(+) progenitor cells ∼225 fold. Herein, we show that using similar technology and subsequent re-expansion methods, we can achieve ∼5 million-fold yields within 24 days of the initial seeding. Interestingly, stem cell phenotype was preserved during the course of the multiple expansions. The high level of the stem cell homing receptor, CXCR4 was expressed in the primary expansion cells, and was maintained throughout the course of re-expansions. In addition, re-expanded cells preserved their multi-potential differential capabilities in vitro, such as, endothelial and smooth muscle lineages. Moreover, biological functionality of the re-expanded cells was preserved and was confirmed by a murine hind limb ischemia model for revascularization. These cells could also be genetically modified for enhanced vasculogenesis. Immunohistochemical evidences support enhanced expression of angiogenic factors responsible for this enhanced neovascularization. These data further confirms that nanofiber-based ex-vivo expansion technology can generate sufficient numbers of biologically functional stem cells for potential clinical applications.
尽管心血管医学领域近来取得了进展，但缺血性疾病仍然是发病和死亡的主要原因。虽然基于干细胞的缺血性疾病治疗方法前景广阔，但具有生物学功能的干细胞供应有限，这阻碍了基于干细胞疗法的应用。此前，我们报道了一种基于聚醚砜（PES）纳米纤维的体外干细胞扩增技术，该技术可使源自人脐带血（UCB）的CD133(+)/CD34(+)祖细胞扩增约225倍。在此，我们表明，使用类似技术及后续再扩增方法，在初始接种后的24天内，我们能够实现约500万倍的产量。有趣的是，在多次扩增过程中干细胞表型得以保留。干细胞归巢受体CXCR-4在初次扩增细胞中高表达，并在再扩增过程中一直保持。此外，再扩增细胞在体外保留了其多能分化能力，如向内皮细胞和平滑肌细胞系分化的能力。而且，再扩增细胞的生物学功能得以保留，并通过小鼠后肢缺血模型的血管再生得到证实。这些细胞还可进行基因改造以增强血管生成。免疫组织化学证据支持了负责这种增强的新血管形成的血管生成因子的表达增强。这些数据进一步证实，基于纳米纤维的体外扩增技术能够产生足够数量的具有生物学功能的干细胞，用于潜在的临床应用。
