Fu H L, Diao Z Y, Shao L, Yang D P
Department of Orthopedic Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China.
Department of Plastic Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China.
Genet Mol Res. 2014 Oct 27;13(4):8620-31. doi: 10.4238/2014.October.27.1.
Osteoporosis poses a major public health threat in aging societies. Adipose-derived stem cells (ADSCs) are multipotent adult stem cells that have the ability to yield mesenchymal stem cells, and have the potential to undergo osteogenesis and bone regeneration. Bone morphogenetic proteins (BMPs) have been demonstrated to upregulate bone gene expression after mechanical injury and to improve bone injury repair. This study aimed to produce BMP-2 expression in ADSCs by using lentiviral vectors. Subcutaneous adipose tissue from 4-week-old male Sprague-Dawley rats was used. Oil red O staining was used to detect adipocyte formation from ADSCs. Induction of ADSC osteogenesis was confirmed with Alizarin red S staining. The recombinant lenti-hBMP-2/neo was constructed to infect ADSCs, BMP-2 expression was measured by immunoblotting analysis, and cellular alkaline phosphatase levels were examined. We found that >70% of ADSC cells could be induced to differentiate into osteocytes or adipocytes. Under osteogenic induction, ADSCs showed increased intracellular calcium deposition, the formation of calcium tubercles, and the disappearance of cellular structures in calcium tubercles. After infection of ADSCs by lenti-hBMP-2/neo, BMP-2 was expressed after doxycycline induction. We, thus, conclude that ADSCs maintain vigorous growth ex vivo and possess stem cell-like properties. When infected with lenti-hBMP-2/neo, ADSCs can be induced to promote BMP-2 expression.
骨质疏松症在老龄化社会中构成了重大的公共卫生威胁。脂肪来源干细胞（ADSCs）是多能成体干细胞，能够产生间充质干细胞，并且具有成骨和骨再生的潜力。骨形态发生蛋白（BMPs）已被证明在机械损伤后可上调骨基因表达，并改善骨损伤修复。本研究旨在通过使用慢病毒载体在ADSCs中产生BMP-2表达。使用4周龄雄性Sprague-Dawley大鼠的皮下脂肪组织。用油红O染色检测ADSCs向脂肪细胞的分化。用茜素红S染色确认ADSC成骨诱导。构建重组慢病毒-hBMP-2/neo感染ADSCs，通过免疫印迹分析测量BMP-2表达，并检测细胞碱性磷酸酶水平。我们发现超过70%的ADSC细胞可被诱导分化为骨细胞或脂肪细胞。在成骨诱导下，ADSCs显示细胞内钙沉积增加、钙结节形成以及钙结节中细胞结构消失。慢病毒-hBMP-2/neo感染ADSCs后，强力霉素诱导后BMP-2表达。因此，我们得出结论，ADSCs在体外保持旺盛生长并具有干细胞样特性。当用慢病毒-hBMP-2/neo感染时，ADSCs可被诱导促进BMP-2表达。
