원문정보
Serum Free Medium Development for Recombinant Erythropoietin Production using Novel Cell Line (QT35)
초록
영어
Human Erythropoietin (EPO) gene is cloned in quail fibrosarcoma cell, QT35. Because molecular weight of EPO is similar to that of serum albumin, cell culture with serum containing medium makes purification of EPO very difficult. Using fractional factorial study, we have developed serum free medium for the recombinant QT35 cell lines, QT N4D4 and QT SY-IMP, which have cloned EPO with glutamine synthetase (GS) gene amplification system and with puromycin selective marker, respectively. Among the seven frequently used medium components, fibronectin, BSA, and EGF were the most important for EPO production. However, sufficient fibronectin supplement to the medium did not make any good attachment of QT35 to culture plate over 3 days. Therefore, to maximize EPO production, we attempted a medium-shift at confluence from serum containing medium to serum free medium(QT SFM6). Using the medium-shift protocol with QT SFM6, nearly the same productivity of EPO was achieved comparing with that without medium-shift. This result was true in both QT35 cell lines in three types of culture, i.e. T flask, microcarrier and roller bottle cultures.
목차
재료 및 방법
세포주(Cell Line)
사용 배지
세포 계대 배양 및 저장
Microcarrier culture 및 roller bottle culture
EPO 역가 분석
결과 및 고찰
무혈청 배지 디자인을 위한 fractional factorial study
QT SFM6 디자인 및 T25 플라스크 배양실험
QT SFM6를 이용한 microcarrier culture
QT SFM6를 이용한 perfusion-type roller bottle culture
요약
감사
참고문헌