Moghadam Farshad Homayouni, Tayebi Tahereh, Dehghan Maryam, Eslami Gilda, Nadri Hamid, Moradi Alireza, Vahedian-Ardakani Hassanali, Barzegar Kazem
Department of Physiology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ; Neurobiomedical Research Center, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Department of Physiology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Int J Hematol Oncol Stem Cell Res. 2014 Oct 1;8(4):12-9.
Bone marrow mesenchymal stem cells (MSCs) are one of the undifferentiated multipotential cell sources of human body. MSCs have the capacity to form a variety of cell types, especially chondrocytes and osteocytes. Learning about responses of MSCs to external milieu and chemical factors such as pH could recommend new approaches for preparation of suitable scaffolds for bone and cartilage tissue engineering. In present study, the effect of alkaline medium on chondrogenic and osteogenic differentiation of rat MSCs was evaluated.
MSCs were harvested from bone marrow of animals and then the response of passage1 and 2 of MSCs (P1 MSCs & P2 MSCs) to the culture in alkaline medium (pH: 8) was evaluated. Cytochemical and immunocytochemical staining were performed to distinguish chondrocytes and osteocytes. Real-time PCR was performed to evaluate the type II collagen and osteopontin mRNA levels.
Staining for type II collagen, a chondrocytic specific marker, revealed that after one-week culture in alkaline medium, a considerable amount of P1 MSCs had shown chondrocytic morphology. By prolonging the culture period up to 4 weeks, osteogenic cells with expanded matrix and mineralized areas around them were appeared. Results of real-time PCR showed that P1 MSCs after one week culture in alkaline medium expressed highest rate of type II collagen and osteopontin mRNA among all groups.
This study demonstrated that alkaline medium is a potent chondrogenic differentiation inducer for MSCs in their first passage.
骨髓间充质干细胞（MSCs）是人体未分化的多能细胞来源之一。MSCs有能力形成多种细胞类型，尤其是软骨细胞和骨细胞。了解MSCs对外部环境和化学因素（如pH值）的反应，可为骨和软骨组织工程制备合适的支架推荐新方法。在本研究中，评估了碱性培养基对大鼠MSCs软骨生成和成骨分化的影响。
从动物骨髓中获取MSCs，然后评估第1代和第2代MSCs（P1 MSCs和P2 MSCs）在碱性培养基（pH值：8）中培养的反应。进行细胞化学和免疫细胞化学染色以区分软骨细胞和骨细胞。进行实时PCR以评估II型胶原蛋白和骨桥蛋白mRNA水平。
对软骨细胞特异性标志物II型胶原蛋白的染色显示，在碱性培养基中培养一周后，相当数量的P1 MSCs呈现出软骨细胞形态。将培养期延长至4周后，出现了基质扩张且周围有矿化区域的成骨细胞。实时PCR结果显示，在碱性培养基中培养一周后的P1 MSCs在所有组中II型胶原蛋白和骨桥蛋白mRNA的表达率最高。
本研究表明，碱性培养基是第一代MSCs的有效软骨生成分化诱导剂。
