A comprehensive protocol for conducting the Syrian hamster embryo cell transformation assay at pH 6.70

被引：41

作者：

Kerckaert, GA ^{[1
]}

Isfort, RJ ^{[1
]}

Carr, GJ ^{[1
]}

Aardema, MJ ^{[1
]}

LeBoeuf, RA ^{[1
]}

机构：

[1] PROCTER & GAMBLE CO, MIAMI VALLEY LABS, HUMAN SAFETY DEPT, CORP P&RS, CINCINNATI, OH 45239 USA

来源：

MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS | 1996年 / 356卷 / 01期

关键词：

Syrian hamster embryo cell; in vitro cell transformation protocol; pH; carcinogen screening;

D O I：

10.1016/0027-5107(95)00198-0

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Studies from our laboratory have demonstrated several advantages of conducting the Syrian hamster embryo (SHE) cell transformation assay at pH 6.70 compared to that done historically at higher pH values (7.10-7.35). These include reduction of the influence of SHE cell isolates and fetal bovine serum lot variability on the assay, an increase in the frequency of chemically induced morphological transformation (MT) compared to controls, and an increased ease in scoring the MT phenotype. The purpose of this paper is to report a comprehensive protocol for conduct of the pH 6.70 SHE transformation assay including experimental procedures, a description of criteria for an acceptable assay and statistical procedures for establishing treatment-related effects. We have also identified several assay parameters in addition to pH which can affect transformation frequencies, particularly the critical role colony number per plate can have on transformation frequency. Control of this parameter, for which details are provided, can greatly increase the reproducibility and predictive value of the assay.

引用

页码：65 / 84

页数：20

共 24 条

[11]

LEBOEUF RA, 1990, PROG CLIN BIOL RES, V340, P219

[12] AN INTERLABORATORY COMPARISON OF ENHANCED MORPHOLOGICAL TRANSFORMATION OF SYRIAN-HAMSTER EMBRYO CELLS CULTURED UNDER CONDITIONS OF REDUCED BICARBONATE CONCENTRATION AND PH [J].

LEBOEUF, RA ;

KERCKAERT, GA ;

POILEY, JA ;

RAINERI, R .

MUTATION RESEARCH, 1989, 222 (03) :205-218

[13]

LEBOEUF RA, 1990, CANCER RES, V50, P3722

[14] ENHANCED MORPHOLOGICAL TRANSFORMATION OF EARLY PASSAGE SYRIAN-HAMSTER EMBRYO CELLS CULTURED IN MEDIUM WITH A REDUCED BICARBONATE CONCENTRATION AND PH [J].

LEBOEUF, RA ;

KERCKAERT, GA .

CARCINOGENESIS, 1987, 8 (05) :689-697

[15]

MCGREGOR D, 1985, PROGR MUTATION RES, V5, P103

[16] MORPHOLOGICAL TRANSFORMATION OF EARLY PASSAGE GOLDEN SYRIAN-HAMSTER EMBRYO CELLS DERIVED FROM CRYOPRESERVED PRIMARY CULTURES AS A RELIABLE INVITRO BIOASSAY FOR IDENTIFYING DIVERSE CARCINOGENS [J].

PIENTA, RJ ;

POILEY, JA ;

LEBHERZ, WB .

INTERNATIONAL JOURNAL OF CANCER, 1977, 19 (05) :642-655

[17] APPLICATION OF TRANSFORMATION SYSTEMS [J].

PIENTA, RJ ;

POILEY, JA ;

RAINERI, R .

ANNALS OF THE NEW YORK ACADEMY OF SCIENCES, 1983, 407 (JUN) :267-283

[18] ROLE OF PLASMA FIBRONECTIN IN THE MORPHOLOGICAL TRANSFORMATION OF HAMSTER-EMBRYO CELLS [J].

RIVEDAL, E .

CANCER LETTERS, 1982, 17 (01) :9-17

[19]

SANNER T, 1985, PROGR MUTATION RES E, V5, P665

[20]

SCHUMAN RF, 1979, IN VITRO CELL DEV B, V15, P730

← 1 2 3 →