碩士 / 國立中興大學 / 化學工程學系所 / 94 / Base on the donors of articular chondrocytes are limited and hard to expand. To obtain adequate chondrocytes and keep acitivity for autologous chondrocytes implantation or tissue engineering research is very important. Expansion by monolayer culture is mostly used. It is well known that chondrocytes cultured with low seeding density and passaged in monolayer will dedifferentiate. Low seeding density can greatly increase the total cell number with less passaging. If a certain number of chondrocytes need to be obtained, light seeding density will dead to frequent passaging. No ideal seeding density and number of passages for chondrocytes in monolayer has been established so far.
In order to understand the effect of cells density and passage in expanding chondrocytes, porcine chondrocytes were expanded in monolayer and in alginate beads. Cell number and the gene expressions were measured.
The study of the gene levels for cells in monolaye culture showed that the initial seeding density of 30000 cells/cm² could maintain the phenotype of chondrocytes. Seeding the dedifferentiated chondrocytes in alginate beads with 1000000 cells/ml could help the fibroblast-like chondrocytes redifferentiate. For chondrocytes from passage one expanded cells from density 5000 cells/cm² in monolayer cultured in alginate beads were monitored higher ECM secretion and chondrogenic gene expression, such as type II collagen and aggrecan gene expression. Chondrocytes in monolayer seeded at a density 5000 cells/cm² expanded faster than at a seeding density 30000 cells/cm² .
Generally, fewer passaging times of chondrocytes with low density in monolayer expansion combinated with higher seeding density in three-dimensional cell carriers are more practical in clinicial applications.
| Identifer | oai:union.ndltd.org:TW/094NCHU5063028 |
| Date | January 2006 |
| Creators | Po-Nien Lin, 林柏年 |
| Contributors | 徐善慧 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 87 |
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