Kilinc Mehmet Okyay, Santidrian Antonio, Minev Ivelina, Toth Robert, Draganov Dobrin, Nguyen Duong, Lander Elliot, Berman Mark, Minev Boris, Szalay Aladar A
Department of Biochemistry, Biocenter, University of Wuerzburg, Am Hubland, 97070, Würzburg, Germany.
StemImmune Inc., San Diego, CA, 92122, USA.
Clin Transl Med. 2018 Feb 7;7(1):5. doi: 10.1186/s40169-018-0183-8.
Stromal vascular fraction (SVF) represents an attractive source of adult stem cells and progenitors, holding great promise for numerous cell therapy approaches. In 2017, it was reported that 1524 patients received autologous SVF following the enzymatic digestion of liposuction fat. The treatment was safe and effective and patients showed significant clinical improvement. In a collaborative study, we analyzed SVF obtained from 58 patients having degenerative, inflammatory, autoimmune diseases, and advanced stage cancer.
Flow analysis showed that freshly isolated SVF was very heterogeneous and harbored four major subsets specific to adipose tissue; CD34 CD45 CD31 CD146 adipose-derived stromal/stem cells (ADSCs), CD34 CD45 CD206CD31 CD146 hematopoietic stem cell-progenitors (HSC-progenitors), CD34 CD45 CD31CD146 adipose tissue-endothelial cells and CD45CD34CD31CD146 pericytes. Culturing and expanding of SVF revealed a homogenous population lacking hematopoietic lineage markers CD45 and CD34, but were positive for CD90, CD73, CD105, and CD44. Flow cytometry sorting of viable individual subpopulations revealed that ADSCs had the capacity to grow in adherent culture. The identity of the expanded cells as mesenchymal stem cells (MSCs) was further confirmed based on their differentiation into adipogenic and osteogenic lineages. To identify the potential factors, which may determine the beneficial outcome of treatment, we followed 44 patients post-SVF treatment. The gender, age, clinical condition, certain SVF-dose and route of injection, did not play a role on the clinical outcome. Interestingly, SVF yield seemed to be affected by patient's characteristic to various extents. Furthermore, the therapy with adipose-derived and expanded-mesenchymal stem cells (ADE-MSCs) on a limited number of patients, did not suggest increased efficacies compared to SVF treatment. Therefore, we tested the hypothesis that a certain combination, rather than individual subset of cells may play a role in determining the treatment efficacy and found that the combination of ADSCs to HSC-progenitor cells can be correlated with overall treatment efficacy.
We found that a 2:1 ratio of ADSCs to HSC-progenitors seems to be the key for a successful cell therapy. These findings open the way to future rational design of new treatment regimens for individuals by adjusting the cell ratio before the treatment.
基质血管成分（SVF）是成体干细胞和祖细胞的一个有吸引力的来源，在众多细胞治疗方法中具有巨大潜力。2017年，有报道称1524例患者在抽脂脂肪经酶消化后接受了自体SVF治疗。该治疗安全有效，患者临床症状有显著改善。在一项合作研究中，我们分析了从58例患有退行性、炎症性、自身免疫性疾病及晚期癌症的患者中获取的SVF。
流式分析显示，新鲜分离的SVF非常异质，包含四个脂肪组织特异性的主要亚群；CD34⁺CD45⁻CD31⁻CD146⁺脂肪来源的基质/干细胞（ADSCs）、CD34⁺CD45⁻CD206⁺CD31⁻CD146⁺造血干细胞祖细胞（HSC-祖细胞）、CD34⁺CD45⁻CD31⁺CD146⁻脂肪组织内皮细胞和CD45⁻CD34⁻CD31⁻CD146⁺周细胞。SVF的培养和扩增显示出一个缺乏造血谱系标志物CD45和CD34但CD90、CD73、CD105和CD44呈阳性的同质群体。对存活的单个亚群进行流式细胞术分选显示，ADSCs有能力在贴壁培养中生长。基于其向脂肪生成和成骨谱系的分化，进一步证实了扩增细胞作为间充质干细胞（MSCs）的身份。为了确定可能决定治疗有益结果的潜在因素，我们在SVF治疗后随访了44例患者。性别、年龄、临床状况、特定的SVF剂量和注射途径对临床结果没有影响。有趣的是，SVF产量似乎在不同程度上受患者特征的影响。此外，对有限数量患者进行的脂肪来源和扩增间充质干细胞（ADE-MSCs）治疗，与SVF治疗相比，并未显示出更高的疗效。因此，我们测试了一种假设，即某种细胞组合而非单个细胞亚群可能在决定治疗效果中起作用，并发现ADSCs与HSC-祖细胞的组合与总体治疗效果相关。
我们发现ADSCs与HSC-祖细胞的2:1比例似乎是细胞治疗成功的关键。这些发现为未来通过在治疗前调整细胞比例为个体合理设计新的治疗方案开辟了道路。
