Department of Oral and Maxillofacial Surgery, The Catholic University of Korea, School of Medicine, Seoul, Korea.
Int J Oral Maxillofac Implants. 2009 Nov-Dec;24(6):991-8.
To investigate the method and conditions of isolation and proliferation of multipotent mesenchymal stem cells (MSCs) from human maxillary sinus membrane in vitro and to induce osteogenic differentiation directly for identification.
A human maxillary sinus membrane specimen was collected in aseptic conditions from an orthognathic surgery patient and cultured. The cells at passage three were sorted by flow cytometry and treated with osteogenic differentiation media. To determine the osteogenic potential of these cells, the authors analyzed alkaline phosphatase (ALP) expression, mineralization of extracellular matrix, and osteocalcin expression; staining with alizarin red and von Kossa and reverse-transcriptase polymerase chain reaction were also performed.
Maxillary sinus membrane-derived cells were positive for STRO-1 and CD105 and negative for CD34. After 7 days, ALP began to be expressed. After 21 and 28 days, most cells showed expression of ALP. Mineralization of the extracellular matrix was observed and, after 21 and 28 days, most of the cells showed mineralization. After 7 days, the osteocalcin gene was expressed; this expression was strongest on the 28th day.
The results suggest that there are MSCs in human maxillary sinus membrane tissue, which can be differentiated into osteoblasts under osteogenic induction. This indicates that maxillary sinus membrane may be a useful source of MSCs for cell therapy.
研究体外分离和扩增人上颌窦黏膜多能间充质干细胞(MSCs)的方法和条件,并直接诱导其向成骨细胞分化进行鉴定。
无菌条件下从正颌手术患者上颌窦膜采集标本进行培养。第 3 代细胞经流式细胞术分选,并用成骨诱导培养基处理。为了确定这些细胞的成骨潜能,作者分析了碱性磷酸酶(ALP)的表达、细胞外基质矿化和骨钙素的表达;茜素红和 von Kossa 染色以及逆转录聚合酶链反应也进行了分析。
上颌窦膜来源的细胞 STRO-1 和 CD105 阳性,CD34 阴性。第 7 天开始表达 ALP。第 21 天和第 28 天,大多数细胞均表达 ALP。观察到细胞外基质的矿化,第 21 天和第 28 天,大多数细胞出现矿化。第 7 天,骨钙素基因开始表达;第 28 天表达最强。
这些结果表明,人上颌窦黏膜组织中存在 MSCs,在成骨诱导下可分化为成骨细胞。这表明上颌窦膜可能是细胞治疗中 MSCs 的一种有用来源。