TY - JOUR
T1 - Clonal insulinoma cell line that stably maintains correct glucose responsiveness
AU - Knaack, David
AU - Fiore, Deborah M.
AU - Surana, Manju
AU - Leiser, Margarita
AU - Laurance, Megan
AU - Fusco-DeMane, David
AU - Hegre, Orion D.
AU - Fleischer, Norman
AU - Efrat, Shimon
PY - 1994
Y1 - 1994
N2 - A number of pancreatic β-tumor cell (βTC) lines have been derived from insulinomas arising in transgenic mice expressing the SV40 T antigen gene under control of the insulin promoter. Some of these lines secrete insulin in response to physiological glucose concentrations. However, this phenotype is unstable. After propagation in culture, these nonclonal lines become responsive to subphysiological glucose levels and/or manifest reduced insulin release. Here we report the use of soft-agar cloning to isolate single-cell clones from a βTC line, which give rise to sublines that maintain correct glucose responsiveness and high insulin production and secretion for >55 passages (over a year) in culture. One of these clonal lines, denoted βTC6- F7, was characterized in detail. βTC6-F7 cells expressed high glucokinase and low hexokinase activity, similarly to normal islets. In addition, they expressed mRNA for the GLUT2 glucose transporter isotype and no detectable GLUT1 mRNA, as is characteristic of normal β-cells. These results demonstrate that transformed β-cells can maintain a highly differentiated phenotype during prolonged propagation in culture, which has implications for the development of continuous β-cell lines for transplantation therapy of diabetes.
AB - A number of pancreatic β-tumor cell (βTC) lines have been derived from insulinomas arising in transgenic mice expressing the SV40 T antigen gene under control of the insulin promoter. Some of these lines secrete insulin in response to physiological glucose concentrations. However, this phenotype is unstable. After propagation in culture, these nonclonal lines become responsive to subphysiological glucose levels and/or manifest reduced insulin release. Here we report the use of soft-agar cloning to isolate single-cell clones from a βTC line, which give rise to sublines that maintain correct glucose responsiveness and high insulin production and secretion for >55 passages (over a year) in culture. One of these clonal lines, denoted βTC6- F7, was characterized in detail. βTC6-F7 cells expressed high glucokinase and low hexokinase activity, similarly to normal islets. In addition, they expressed mRNA for the GLUT2 glucose transporter isotype and no detectable GLUT1 mRNA, as is characteristic of normal β-cells. These results demonstrate that transformed β-cells can maintain a highly differentiated phenotype during prolonged propagation in culture, which has implications for the development of continuous β-cell lines for transplantation therapy of diabetes.
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U2 - 10.2337/diab.43.12.1413
DO - 10.2337/diab.43.12.1413
M3 - Article
C2 - 7958492
AN - SCOPUS:0028577616
SN - 0012-1797
VL - 43
SP - 1413
EP - 1417
JO - Diabetes
JF - Diabetes
IS - 12
ER -