Y Jiao1, X Ma, S Yu, M Shao. 1. School of Stomatology, Beijing Medical University.
Abstract
OBJECTIVE: The purpose of this study was to investigate the effects of exogenous fibronectin on the attachment and growth of mandibular condylar cartilage (MCC) cells on DEAE-dextran micro-carrier. METHODS: MCC cells were harvested from newborn New Zealand white rabbits by sequential digestion with trypsin and collagenase. They were grown on the 20 mg/L fibronectin-coated and uncoated cytodex-3 micro-carrier. Samples were collected after 1, 3, 5, 7, 9, 11 days. The kinetics of adhesion and growth were observed with phase contrast microscope and environmental scanning electronic microscope (ESEM) and quantified by 0.1% crystal violet nuclear extrusion method. RESULTS: MCC cells could rapidly attach and spread onto the fibronectin-coated cytodex-3, and nearly 60 percent cells attached to the micro-carrier within 2 hours. Spreading cells were flat and lowly refractile observed under phase contrast microscope. While the attachment and spreading of the cells in the untreated group were slow, and only 36 percent cells attached to the uncoated micro-carrier. However, most cells attached to the micro-carrier in both groups after 24 hours. The MCC cells attaching to FN-coated cytodex-3 observed under ESEM were flatter and adhered tightly with more processes (pseudopodia) stretching out from cytoplasm. In contrast, cells on untreated cytodex-3 remained regularly round even 24 hours after being plated. Meanwhile, there was an accelerated growth rate of the MCC cells on the fibronectin treated cytodex-3. The cell density in the FN treated group was higher than that of the untreated group at the end of the culture. CONCLUSION: Fibronectin, one major glycoprotein of extracellular matrix, plays an important role in cell attachment and spreading. The better understanding of the extracellular matrix will be helpful in selecting the optimal substrate for cell growth in vitro.
OBJECTIVE: The purpose of this study was to investigate the effects of exogenous fibronectin on the attachment and growth of mandibular condylar cartilage (MCC) cells on DEAE-dextran micro-carrier. METHODS: MCC cells were harvested from newborn New Zealand white rabbits by sequential digestion with trypsin and collagenase. They were grown on the 20 mg/L fibronectin-coated and uncoated cytodex-3 micro-carrier. Samples were collected after 1, 3, 5, 7, 9, 11 days. The kinetics of adhesion and growth were observed with phase contrast microscope and environmental scanning electronic microscope (ESEM) and quantified by 0.1% crystal violet nuclear extrusion method. RESULTS: MCC cells could rapidly attach and spread onto the fibronectin-coated cytodex-3, and nearly 60 percent cells attached to the micro-carrier within 2 hours. Spreading cells were flat and lowly refractile observed under phase contrast microscope. While the attachment and spreading of the cells in the untreated group were slow, and only 36 percent cells attached to the uncoated micro-carrier. However, most cells attached to the micro-carrier in both groups after 24 hours. The MCC cells attaching to FN-coated cytodex-3 observed under ESEM were flatter and adhered tightly with more processes (pseudopodia) stretching out from cytoplasm. In contrast, cells on untreated cytodex-3 remained regularly round even 24 hours after being plated. Meanwhile, there was an accelerated growth rate of the MCC cells on the fibronectin treated cytodex-3. The cell density in the FN treated group was higher than that of the untreated group at the end of the culture. CONCLUSION: Fibronectin, one major glycoprotein of extracellular matrix, plays an important role in cell attachment and spreading. The better understanding of the extracellular matrix will be helpful in selecting the optimal substrate for cell growth in vitro.