F9 기형암종 세포의 미세전이 Assay 방법의 개발
Development of an Assay System for the Micrometastasis of F9 Teratocarcinoma Stem Cell
대한암학회지 1994년 26권 2호 p.304 ~ 310
차희재, 김철우, 박병채, 김규원,
소속 상세정보
차희재 ( )
부산대학교
김철우 ( )
부산대학교
박병채 ( )
부산대학교
김규원 ( )
부산대학교
KMID : 0360319940260020304
Abstract
During the tumor progression, micrometastases at their earliest stages have been difficult to analyze qualitatively or quantitatively because of lacking suitable sensitive markers to discriminate small numbers of tumor cells from cell populations
of
normal tissue. To identify the distribution of micrometastases and tracing of F9 teratocarcinoma stem cell in vivo, we used chick embryos and the polymerase china reaction(PCR) as simple, sensitive, and quantitative assay system. Hence, F9 cells
which
metastasized in the liver of chick embryo were detected by PCR using specific primers for mouse β-globin gene.
In addition, another assay method was developed for detecting micrometastases of F9 cells with Ecoli lacZ gene as a genetical and histochemical marker. After the subsequent transfection of the lacZ gene into F9 cells, an intensely blue-staining
clone
harboring lacZ gene was isolated by using chromogenic substrate, 5-bromo-4-chloro-3-indoly-β-D-galacto-pyranoside (X-Gal). The lacZ-bearing F9 cells at primary tumor sites as well as at secondary organs can be stained intensely blue spots and
easily
distinguished from the host tissue cells after hours, days, or weeks postinjection.
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