Department of Bio-medical Sciences, University of Catania, Catania, Italy.
Exp Biol Med (Maywood). 2011 Nov;236(11):1333-41. doi: 10.1258/ebm.2011.011183. Epub 2011 Oct 28.
The present study focused on the isolation, cultivation and characterization of human mesenchymal stem cells (MSCs) from adipose tissue and on their differentiation into chondrocytes through the NH ChondroDiff medium. The main aim was to investigate some markers of biomechanical quality of cartilage, such as lubricin, and collagen type I and II. Little is known, in fact, about the ability of chondrocytes from human MSCs of adipose tissue to generate lubricin in three-dimensional (3D) culture. Lubricin, a 227.5-kDa mucinous glycoprotein, is known to play an important role in articular joint physiology, and the loss of accumulation of lubricin is thought to play a role in the pathology of osteoarthritis. Adipose tissue is an alternative source for the isolation of multipotent MSCs, which allows them to be obtained by a less invasive method and in larger quantities than from other sources. These cells can be isolated from cosmetic liposuctions in large numbers and easily grown under standard tissue culture conditions. 3D chondrocytes were assessed by histology (hematoxylin and eosin) and histochemistry (Alcian blue and Safranin-O/fast green staining). Collagen type I, II and lubricin expression was determined through immunohistochemistry and Western blot. The results showed that, compared with control cartilage and monolayer chondrocytes showing just collagen type I, chondrocytes from MSCs (CD44-, CD90- and CD105- positive; CD45-, CD14- and CD34-negative) of adipose tissue grown in nodules were able to express lubricin, and collagen type I and II, indicative of hyaline cartilage formation. Based on the function of lubricin in the joint cavity and disease and as a potential therapeutic agent, our results suggest that MSCs from adipose tissue are a promising cell source for tissue engineering of cartilage. Our results suggest that chondrocyte nodules producing lubricin could be a novel biotherapeutic approach for the treatment of cartilage abnormalities.
本研究专注于从脂肪组织中分离、培养和鉴定人骨髓间充质干细胞（MSCs），并通过 NH 软骨分化培养基将其分化为软骨细胞。主要目的是研究一些软骨生物力学质量的标志物，如润滑素、I 型和 II 型胶原。事实上，人们对脂肪组织来源的人 MSC 软骨细胞在三维（3D）培养中产生润滑素的能力知之甚少。润滑素是一种 227.5kDa 的黏蛋白糖蛋白，已知在关节生理中发挥重要作用，而润滑素的积累减少被认为在骨关节炎的病理学中起作用。脂肪组织是分离多能 MSC 的替代来源，它可以通过一种微创方法从脂肪组织中获得大量的 MSC，比从其他来源获得的 MSC 更多。这些细胞可以从美容吸脂中大量分离出来，并在标准组织培养条件下轻松生长。3D 软骨细胞通过组织学（苏木精和伊红）和组织化学（阿尔辛蓝和番红 O/固绿染色）进行评估。通过免疫组织化学和 Western blot 检测 I 型、II 型胶原和润滑素的表达。结果表明，与对照软骨和单层软骨细胞仅表达 I 型胶原相比，在结节中生长的脂肪组织来源的 MSC（CD44-、CD90-和 CD105-阳性；CD45-、CD14-和 CD34-阴性）的软骨细胞能够表达润滑素和 I 型、II 型胶原，表明形成透明软骨。基于润滑素在关节腔中的功能和疾病以及作为潜在的治疗剂，我们的结果表明，脂肪组织来源的 MSC 是软骨组织工程的有前途的细胞来源。我们的结果表明，产生润滑素的软骨细胞结节可能是治疗软骨异常的一种新的生物治疗方法。
