Dykstra Jordan A, Blue Elliot D, Negrão de Assis Pedro L, Weimer Jill M, Kota Daniel Jiro
Sanford School of Medicine, University of South Dakota, Vermillion, SD 57069, USA.
Sanford Research, Sioux Falls, SD 57104, USA: These authors contributed equally.
J Stem Cells Regen Med. 2020 May 27;16(1):16-25. doi: 10.46582/jsrm.1601004. eCollection 2020.
Due to their capacity to self-renew, proliferate and generate multi-lineage cells, adult-derived stem cells offer great potential in regenerative therapies to treat maladies such as diabetes, cardiac disease, neurological disorders and orthopedic injuries. Commonly derived from adipose tissue, the stromal vascular fraction (SVF), a heterogeneous cell population enriched with mesenchymal stem cells (MSCs), has garnered interest as a cellular therapy due to ease of accessibility as an autologous, point-of-care application. However, the heterogeneous cell population within SVF is not historically taken into consideration when injecting into patients. Here, we characterized SVF, determined its safety and verify its therapeutic effects in a NOD/scid mouse model of articular injury. SVF were isolated from lipoaspirates utilizing a commercially available system (InGeneron Inc.), while MSCs were isolated from SVF via cell culture. Flow cytometry showed that neither age nor BMI affects the frequency of progenitor cells-like (CD31+CD34+), immune cells-like (CD4+) T cells, (CD14+) monocytes and total number of cells obtained. However, there was a negative correlation between donor BMI and MSC frequency within the SVF. ELISAs showed that following LPS activation in SVF, there were low levels of TNF-α and high levels of IL-10 secreted. However, T cell activation with anti-CD3 or anti-CD3+ anti-CD28, while leading to expected high levels of IFN-γ, did not lead to significant levels of TGF-β. PCR analysis showed no significant numbers of cells outside the joint 1-hour post injection, moreover, no engraftment or abnormal growth in other organs 60-days post injection. Finally, both cell populations were able to ameliorate disease progression, as confirmed by the increase in movement of treated groups compared to injured groups. Noteworthy, the histological analysis indicated that there was no cartilage growth, suggesting an alternative therapeutic mechanism to cartilage regeneration.
由于成体干细胞具有自我更新、增殖并生成多谱系细胞的能力，其在再生疗法中具有巨大潜力，可用于治疗糖尿病、心脏病、神经紊乱和骨科损伤等疾病。基质血管成分（SVF）通常源自脂肪组织，是一个富含间充质干细胞（MSC）的异质性细胞群体，因其作为自体即时护理应用易于获取，已成为一种细胞疗法并引起了人们的关注。然而，在向患者注射时，历史上并未考虑SVF中的异质性细胞群体。在此，我们对SVF进行了表征，确定了其安全性，并在关节损伤的NOD/scid小鼠模型中验证了其治疗效果。使用市售系统（InGeneron公司）从抽脂物中分离SVF，而通过细胞培养从SVF中分离MSC。流式细胞术显示，年龄和体重指数（BMI）均不影响祖细胞样（CD31+CD34+）、免疫细胞样（CD4+）T细胞、（CD14+）单核细胞的频率以及获得的细胞总数。然而，供体BMI与SVF内MSC频率之间存在负相关。酶联免疫吸附测定（ELISA）表明，在SVF中进行脂多糖（LPS）激活后，分泌的肿瘤坏死因子-α（TNF-α）水平较低，白细胞介素-10（IL-10）水平较高。然而，用抗CD3或抗CD3 + 抗CD28激活T细胞，虽然导致预期的高水平干扰素-γ（IFN-γ），但并未导致显著水平的转化生长因子-β（TGF-β）。聚合酶链反应（PCR）分析显示，注射后1小时关节外无大量细胞，此外，注射后60天其他器官无植入或异常生长。最后，与损伤组相比，治疗组运动增加证实了这两种细胞群体均能够改善疾病进展。值得注意的是，组织学分析表明没有软骨生长，提示存在一种不同于软骨再生的治疗机制。
