Enhancement of Cell Growth on Microcarriers Immobilized with Insulin
Other Title
微載體表面之胰島素固定化處理對細胞生長之促進
Journal
Journal of the Chinese Institute of Chemical Engineers
Journal Volume
32
Journal Issue
2
Start Page
125
End Page
133
ISSN
0368-1653
Date Issued
2001-03
Author(s)
Abstract
Mammalian cell culture technology has been used to produce structurally complex protein molecules and to construct devices for tissue engineering applications. This study investigated the cultivation of Chinese hamster ovary (CHO) cells on Cytodex 1 microcarriers in a spinner flask. The initial cell attachment was rapid with a rate constant of 5.56×10-2min-1under the condition of continuous stirring at 40 rpm using 2 g l-1 of the microcarriers in a DMEM/F12 (1:1) medium. Adding serum to the medium decreased the rate of cell attachment in a dose-dependent manner. In contrast, the cell growth rate and maximum cell density increased with a shorter lag phase as the concentration of serum in the medium rose. To reduce the cost of the cell culture, the surfaces of Cytodex 1 microcarriers were immobilized with insulin using an adsorption-drying method and then used for the cultivation of CHO cells in a serum-free medium. The growth rat of CHO cells on insulin-immobilized microcarriers using a serum-free medium was comparable to that on untreated microcarriers using a 2% serum-containing medium. In addition, insulin immobilization also enhanced the value of the growth yield for glucose (Ycell/glucose) from 2.41×105 to 3.66×105 cells μmole-1. This finding suggests that the immobilized insulin promoted glucose utilization, thereby enhancing cell growth. The immobilization of insulin on microcarrier is, thus, highly recommended to improve the outcome achieved in a serum-free culture.
動物細胞培養技術已被用於生產結構複雜的蛋白質分子，目前更進一步用於建構組織工程應用所需各項裝置。本研究以旋轉瓶為生物反應器，探討中國大頰鼠卵巢（Chinese hamster ovary, CHO）細胞在Cytodex 1微戴體表面培養之條件。採用40rpm連續攪拌以及微載體濃度為2 g/l的DMEM/F12(1:1)無血清培養液可使細胞在接種後達到快速附著的效果（速率常數為5.56×10-2min-1）。研究若培養液中添加血清則細胞的附著速率會隨著血清濃度的提高而下降，但細胞生長速率以及細胞最高密度會隨著血清濃度提高而增加。為降低細胞培養成本，本研究將胰島表以吸附-乾燥法固定於微載體表面，配合無血清培養液用於培養CHO細胞。結果顯示在上述條件下細胞生長速率能夠接近含2%血清培養液所獲致的生長效果。胰島素固定於微載體表面可使葡萄糖的細胞生長產率（Ycell/glucose）由2.41×105 to 3.66×105 cells μmole-1。此數據說明被固定的胰島素能促進葡萄糖利用並藉此增進細胞生長。本文因而推薦採用固定有胰島素的微載體以提升無血清細胞培養的成效。
Subjects
CHO Cells
Immobilization
Insulin
Microcarriers
Type
journal article