Evaluation of the alginate culture system for chondrogenesis of human adipose tissue-derived stromal cells and assessment of the rotational speed for the novel bioreactor vessel / 褐藻膠培養系統對人類脂肪幹細胞軟骨新生之評估及新型生物反應器動態培養條件之評估

碩士 / 國立中興大學 / 化學工程學系 / 93 / In view of that the three-dimensional (3-D) structure was necessary for chondrogenesis, we fabricated alginate beads and pads to be the cell carriers. Human adipose tissue derived stromal cell (ADSC) were embedded in the gel to study the effects of inducers, culture model and components of extracellular matrix on chondrogenesis. Transforming growth factor-beta 3 (TGF-β3) was found to be a better inducer for chondrogenesis of ADSC than TGF-β1. Cell proliferation was limited in divalent cations crosslinked alginate beads; however, matrix synthesis and deposition were benefited from the 3-D environment. Addition of hyaluronic acid in alginate gel increased type II collagen and GAG contents, while addition of chondroitin sulfate promoted cell proliferation and up-regulated the aggrecan gene expression. In the aspect of bioreactor, better results of cell number, scaffold dry weight, mechanical strength and total GAG contents were obtained from dynamic culture than static culture. At the same time, dynamic culture condition stimulated the cells to secrete NO, which could reduce collagen synthesis. Large amount of GAG was released to media upon the flush arising from the rotation. Based on the data of cell proliferation, matrix accumulation and mechanical strength, the optimal rotational speed for the novel bioreactor vessel was smaller than 25 rpm. This study provided a carrier system for chondrogenesis and established suitable parameters for the bioreactor.

Identiferoai:union.ndltd.org:TW/093NCHU0063048
Date January 2005
CreatorsYing-Ying-Yi Lee, 李盈誼
ContributorsShan-hui Hsu, 徐善慧
Source SetsNational Digital Library of Theses and Dissertations in Taiwan
Languagezh-TW
Detected LanguageEnglish
Type學位論文 ; thesis
Format103

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