Institute of Cellular Medicine, Faculty of Medical Sciences, Newcastle University, International Centre for Life, Times Square, Newcastle-upon-Tyne, NE1 4EP, UK.
Stem Cell Rev Rep. 2013 Oct;9(5):599-608. doi: 10.1007/s12015-013-9446-3.
Satisfactory osseous tissue integration of the soft tissue graft with bone is the mainstay of healing following surgical reconstruction of the anterior cruciate ligament (ACL). However, tissue remodelling is slow and significantly impacts on quality of life by delaying return to work and sport and accelerating the onset of degenerative diseases such as osteoarthritis. Delivery of multipotent human mesenchymal stem cells (hMSCs) at surgery could enhance osseous tissue integration. We aim to use hMSCs derived from haemarthrosis fluid (HF) (the intra-articular bleed accrued post-trauma) which is aspirated and discarded as clinical waste. With the aim of improving our bioprocessing methodologies for clinical translation we have investigated the effect of low oxygen tension on the derivation and osteogenic potential of this novel HF-hMSC population. Mononuclear cells were isolated from HF aspirated samples and divided for derivation and culture under normal or low oxygen tension. HF-hMSCs were derived from 100 % of cultures under low oxygen tension compared to 71 % for normal oxygen tension; this was coupled with increased CFU-Fs. We investigated the osteogenic potential and cellular health of HF-hMSC populations following ex vivo expansion. HF-hMSC populations showed enhanced matrix mineralisation and cellular health when differentiated under low oxygen tension. This positive effect of low oxygen on osteogenesis and cellular health was reduced with prolonged culture. These data demonstrate that derivation and culture of HF-hMSC populations under low oxygen tension will enable the translation of a cellular therapy for the treatment of broad patient numbers with optimal osteogenic potency and cellular vitality.
软组织移植物与骨组织的令人满意的骨组织整合是前交叉韧带（ACL）重建术后愈合的主要支柱。然而，组织重塑缓慢，通过延迟重返工作和运动以及加速退行性疾病（如骨关节炎）的发生，对生活质量有重大影响。在手术时递送多能人骨髓间充质干细胞（hMSC）可以增强骨组织整合。我们旨在利用源自关节内积血（HF）（创伤后关节内积累的出血）的 hMSC，HF 是作为临床废物被抽吸和丢弃的。为了改进我们的临床转化生物处理方法，我们研究了低氧张力对这种新型 HF-hMSC 群体的衍生和成骨潜能的影响。从 HF 抽吸样本中分离出单核细胞，并在正常或低氧张力下进行衍生和培养。与正常氧张力相比，低氧张力下可从 100%的培养物中衍生出 HF-hMSC，这与 CFU-Fs 的增加有关。我们研究了 HF-hMSC 群体在体外扩增后的成骨潜能和细胞健康状况。在低氧张力下分化时，HF-hMSC 群体显示出增强的基质矿化和细胞健康。低氧对成骨作用和细胞健康的这种积极影响随着培养时间的延长而降低。这些数据表明，在低氧张力下衍生和培养 HF-hMSC 群体将能够转化细胞疗法，以最佳的成骨效力和细胞活力治疗广泛的患者群体。
