Functional analysis of a conditionally transformed pancreatic beta-cell line.

Fleischer, Norman; Chen, Chuan; Surana, Manju; Leiser, Margarita; Rossetti, Luciano; Pralong, William; Efrat, Shimon

doi:10.2337/diabetes.47.9.1419

Cited by 89 publications

(95 citation statements)

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“…This long-term exposure to high glucose concentrations does not seem to result in glucose toxicity, as judged by the normal insulin production and glucose sensing maintained in βTC-tet cells for over 60 passages [15]. However, these culture conditions might have been expected to confuse the proliferation assay, since the glucose concentration used could itself have a stimulatory effect on cell replication.…”

Section: Resultsmentioning

confidence: 99%

Growth Factor‐Dependent Proliferation of the Pancreatic β‐cell Line βTC‐tet: An Assay for β‐cell Mitogenic Factors

Milo-Landesman

Efrat

2001

Journal of Diabetes Research

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The ability to expand normal pancreatic islet β cells in culture would significantly advance the prospects of cell therapy for diabetes. A number of growth factors can stimulate limited islet cell replication, however other factors may exist which are more effective β-cell-specific mitogens. The search for novel β-cell growth factors has been hampered by the lack of a β-cell-specific proliferation assay. We developed a simple and sensitive assay for β-cell growth factors based on a conditionally-transformed mouse β-cell line (βTC-tet). These cells express the SV40 T antigen (Tag) oncoprotein under control of the tetracycline (Tc) operon regulatory system. In the presence of Tc, Tag expression is tightly shut off and the cells undergo complete growth arrest. Here we show that the growth-arrested cells can proliferate in response to growth factors in the absence of Tag. Using this assay, a number of growth factors previously shown to be mitogenic to a mixed islet cell population were found to induce proliferation of pure β cells. We conclude that growth-arrested βTC-tet cells can be employed in a survey of factors from various sources for identifying novel factors with β-cell mitogenic activity.Key words: β-cell lines, cell proliferation, growth factors, mitogenicity, tetracycline-regulated gene expression INTRODUCTIONBeta-cell transplantation represents an attractive approach for replacement of the ____________________ *Corresponding author: tel: 972-3-640 7701; fax. 972-3-640 9950; e-mail: sefrat@post.tau.ac.il Int. Jnl. Experimental Diab. Res., 3:69-74, 2002 © 2002 Taylor and Francis 1560 damaged β cells in type I diabetes. One major obstacle to β-cell therapy is the lack of an abundant cell supply. A number of rodent β-cell lines have been developed by oncogenic transformation, however this approach has proven difficult to adapt to human β cells. Development of efficient ways for in vitro expansion of normal islets would significantly advance the prospects of diabetes cell therapy. Adult islet cells are difficult to propagate in culture, although they maintain a proliferative capacity, as evidenced by a limited mitogenic response to a number of growth factors [1,2]. Members of the growth hormone family, in particular placental lactogen (PL) and prolactin (PRL), induce replication in neonatal rat islet cells [3]. These activities may be responsible for islet mass expansion in pregnancy. Significant mitogenic effects of hepatocyte growth factor (HGF) have been observed on human fetal and adult islets [4] and mouse islets [5]. Insulin-like growth factors (IGF) I and II, and platelet-derived growth factor, affect fetal rat islet cell growth [6,7]. IGF I has also been shown to stimulate replication of cultured rat insulinoma cells [8], and IGF II activates the growth of both normal [9] and transformed [10] mouse β cells in vivo. The Reg protein, produced by pancreatic acinar cells and regenerating islets, is another candidate for a β-cell growth factor [11]. These findings suggest that islet cells possess cell su...

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Section: Resultsmentioning

confidence: 99%

Growth Factor‐Dependent Proliferation of the Pancreatic β‐cell Line βTC‐tet: An Assay for β‐cell Mitogenic Factors

Milo-Landesman

Efrat

2001

Journal of Diabetes Research

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“…Insulin secretion from FH-B-TPN cells was measured by static incubation as described (2). Cells were plated in 24-well plates at 10 5 cells per well.…”

Section: Methodsmentioning

confidence: 99%

“…An effective cell replacement strategy depends on the development of an abundant supply of beta cells and their protection from immune destruction. Reversible immortalization of differentiated beta cells by conditional oncogene expression allowed the controlled expansion of murine cells in tissue culture, which were capable of replacing beta-cell function in vivo (1)(2)(3). However, this approach has not been successful yet with beta cells from human islets, which tend to lose insulin expression during forced replication (4).…”

mentioning

confidence: 99%

Reversal of hyperglycemia in mice by using human expandable insulin-producing cells differentiated from fetal liver progenitor cells

Zalzman

Gupta

Giri

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

246

148

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Beta-cell replacement is considered to be the most promising approach for treatment of type 1 diabetes. Its application on a large scale is hindered by a shortage of cells for transplantation. Activation of insulin expression, storage, and regulated secretion in stem͞progen-itor cells offers novel ways to overcome this shortage. We explored whether fetal human progenitor liver cells (FH) could be induced to differentiate into insulin-producing cells after expression of the pancreatic duodenal homeobox 1 (Pdx1) gene, which is a key regulator of pancreatic development and insulin expression in beta cells. FH cells possess a considerable replication capacity, and this was further extended by introduction of the gene for the catalytic subunit of human telomerase. Immortalized FH cells expressing Pdx1 activated multiple beta-cell genes, produced and stored considerable amounts of insulin, and released insulin in a regulated manner in response to glucose. When transplanted into hyperglycemic immunodeficient mice, the cells restored and maintained euglycemia for prolonged periods. Quantitation of human C-peptide in the mouse serum confirmed that the glycemia was normalized by the transplanted human cells. This approach offers the potential of a novel source of cells for transplantation into patients with type 1 diabetes.

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“…Also, this approach carries with it the possibility of oncogenic transformation. [162][163][164][165] Although still controversial, there are data indicating that mature human b cells can be induced to replicate under the effects of hepatocyte growth factor (HGF). [166][167][168] The limitation of this approach, however, rests on the loss of differentiation of the induced b-cell along with a substantial decrease in insulin production.…”

Section: Stem/progenitor Cellsmentioning

confidence: 99%