A protocol to enhance INS1E and MIN6 functionality—The use of theophylline

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Abstract

In vitro research in the field of type I diabetes is frequently limited by the availability of a functional model for islets of Langerhans. This method shows that by the addition of theophylline to the glucose buffers, mouse insulinoma MIN6 and rat insulinoma INS1E pseudo-islets can serve as a model for islets of Langerhans for in vitro research. The effect of theophylline is dose- and cell line-dependent, resulting in a minimal stimulation index of five followed by a rapid return to baseline insulin secretion by reducing glucose concentrations after a first high glucose stimulation. This protocol solves issues concerning in vitro research for type I diabetes as donors and the availability of primary islets of Langerhans are limited. To avoid the limitations of using human donor material, cell lines represent a valid alternative. Many different β cell lines have been reported, but the lack of reproducible responsiveness to glucose stimulation remains a challenge.

Figures

  • Figure 1. The effect of theophylline on insulin secretion. Insulin secretion (µg/L) (A) and stimulation index (B) of MIN6 cells cultured on tissue culture plastic (monolayer) without theophylline, with theophylline 10 mM in all the incubation buffers (T), and with theophylline 10 mM added only in the high glucose buffer (T(hg)).
  • Figure 2. The effect of theophylline on the metabolic activity of MIN6 and INS1E cells. Metabolic activity of MIN6 (A) or INS1E (B) pseudo-islets cultured for 5 h in presence of different concentrations of theophylline. Data are presented as percentage of the metabolic activity at t = 0.
  • Figure 3. Theophylline concentration-dependent insulin secretion of MIN6 pseudo-islets. Doseresponse results of the glucose-induced insulin secretion test on MIN6 pseudo-islets at different concentrations of theophylline. (A) Secreted amount of insulin (µg/L) is shown; in (B) data are normalized by the amount of insulin secreted in low glucose condition (stimulation index). Data are expressed as mean ± standard deviation and significant differences are indicated with * (p ≤ 0.05). Statistical analysis compares the stimulation indices in high glucose conditions.
  • Figure 3. Theophylline concentration-dependent insulin secretion of MIN6 pseudo-islets. Doseresponse results of the glucose-induced insulin secretion test on MIN6 pseudo-islets at different concentrations of theophylline. (A) Secreted amount of insulin (µg/L) is shown; in (B) data are normalized by the amount of insulin secreted in low glucose condition (stimulation index). Data are expressed as mean ± standard deviation and significant differences are indicated with * (p ≤ 0.05). Statistical analysis compares the stimulation indices in high glucose conditions.
  • Figure 4. Theophylline concentration–dependent insulin secretion of INS1E pseudo-islets. Doseresponse results of the glucose-induced insulin secretion test of INS1E pseudo-islets at different concentrations of theophylline. (A) Secreted amount of insulin (µg/L) is shown; (B) Data are normalized by the amount of insulin secreted in low glucose condition (stimulation index). Data are expressed as mean ± standard deviation and significant differences are indicated with * (p ≤ 0.05).

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CITATION STYLE

APA

Nibbelink, M. G., Marchioli, G., Moroni, L., Karperien, M., & Van Apeldoorn, A. (2016). A protocol to enhance INS1E and MIN6 functionality—The use of theophylline. International Journal of Molecular Sciences, 17(9). https://doi.org/10.3390/ijms17091532

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