Chun So Young, Kim Hyun Tae, Kwon Se Yun, Kim Jeongshik, Kim Bum Soo, Yoo Eun Sang, Kwon Tae Gyun
1Bio Medical Research Institute, Kyungpook National University Hospital, Daegu, Korea.
2Department of Urology, School of Medicine, Kyungpook National University, Daegu, Korea.
Tissue Eng Regen Med. 2016 Aug 5;13(4):403-415. doi: 10.1007/s13770-016-9073-6. eCollection 2016 Aug.
Upper urinary tract-derived urine stem cells (USCs) are considered a valuable mesenchymal stem cell source for autologous cell therapy. However, the reported culture condition for USCs is not appropriate for large-quantity production, because cells can show limited replicativity, senescence, and undesirable differentiation during cultivation. These drawbacks led us to reconstitute a culture condition that mimics the natural stem cell niche. We selected extracellular matrix protein and oxygen tension to optimize the expansion of USCs, and compared cell adhesion, proliferation, gene expression, chromosomal stability, differentiation capacity, immunity and safety. Culture on collagen type I (ColI) supported highly enhanced USC proliferation and retention of stem cell properties. In the oxygen tension analysis (with ColI), 5% O hypoxia showed a higher cell proliferation rate, a greater proportion of cells in the S phase of the cell cycle, and normal stem cell properties compared to those observed in cells cultured under 20% O normoxia. The established reconstituted condition (ColI/hypoxia, USCs) was compared to the control condition. The expanded USCs showed highly increased cell proliferation and colony forming ability, maintained transcription factors, chromosomal stability, and multi-lineage differentiation capacity (neuron, osteoblast, and adipocyte) compared to the control. In addition, USCs retained their immune-privileged potential and non-tumorigenicity with testing at week 8. Therefore, reconstituted condition allows for expanded uUSC cell preparations that are safe and useful for application in stem cell therapy.
上尿路来源的尿液干细胞（USCs）被认为是自体细胞治疗中一种有价值的间充质干细胞来源。然而，已报道的USCs培养条件并不适合大量生产，因为在培养过程中细胞的增殖能力有限、会出现衰老以及发生不良分化。这些缺点促使我们重新构建一种模拟天然干细胞微环境的培养条件。我们选择细胞外基质蛋白和氧张力来优化USCs的扩增，并比较细胞黏附、增殖、基因表达、染色体稳定性、分化能力、免疫性和安全性。在I型胶原（ColI）上培养可高度增强USCs的增殖并保留干细胞特性。在氧张力分析（使用ColI）中，与在20%氧常氧条件下培养的细胞相比，5%氧低氧条件下细胞增殖率更高，处于细胞周期S期的细胞比例更大，且具有正常的干细胞特性。将建立的重组培养条件（ColI/低氧，USCs）与对照条件进行比较。与对照组相比，扩增后的USCs细胞增殖和集落形成能力显著增强，维持了转录因子、染色体稳定性以及多向分化能力（神经元、成骨细胞和脂肪细胞）。此外，在第8周进行检测时，USCs保留了其免疫特权潜力和非致瘤性。因此，重组培养条件能够扩增出安全且可用于干细胞治疗的尿源性USCs细胞制剂。
