Li Yong, Pan Haiying, Huard Johnny
Stem Cell Research Center, Childrens Hospital of Pittsburgh of UPMC, PA, USA.
J Vis Exp. 2010 Jul 5(41):2011. doi: 10.3791/2011.
Adult stem cells have long been discussed in regards to their application in regenerative medicine. Adult stem cells have generated a great deal of excitement for treating injured and diseased tissues due to their impressive capabilities to undergo multi-lineage cell differentiation and their self-renewal ability. Most importantly, these qualities have made them advantageous for use in autologous cell transplantation therapies. The current protocol will introduce the readers to the modified preplate technique where soft tissues of the musculoskeletal system, e.g. tendon and muscle, are 1(st) enzymatically dissociated and then placed in collagen coated flasks with medium. The supernatant, which is composed of medium and the remaining floating cells, is serially transferred daily to new flasks. The stem cells are the slowest to adhere to the flasks which is usually takes 5-7 days (serial transfers or preplates). By using this technique, adult stem cells present in these tissues can be easily harvested through fairly non-invasive procedures.
长期以来,人们一直在讨论成体干细胞在再生医学中的应用。成体干细胞因其令人印象深刻的多谱系细胞分化能力和自我更新能力,在治疗受损和患病组织方面引起了极大的关注。最重要的是,这些特性使其在自体细胞移植治疗中具有优势。本方案将向读者介绍改良的预铺板技术,即对肌肉骨骼系统的软组织(如肌腱和肌肉)进行酶解,然后置于涂有胶原蛋白的培养瓶中,并加入培养基。由培养基和剩余悬浮细胞组成的上清液每天依次转移到新的培养瓶中。干细胞附着到培养瓶的速度最慢,通常需要5 - 7天(连续转移或预铺板)。通过使用这种技术,可以通过相当非侵入性的程序轻松收获这些组织中存在的成体干细胞。