Novel porous matrix and bioreactors for high density cultures of insulinoma cell lines: Insulin secretion and response to glucose
✍ Scribed by Anita Handa-Corrigan; Sima Hayavi; Hazem Ghebeh; Nesredin A. Mussa; Marion Chadd
- Publisher
- Wiley (John Wiley & Sons)
- Year
- 1998
- Tongue
- English
- Weight
- 390 KB
- Volume
- 71
- Category
- Article
- ISSN
- 0268-2575
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✦ Synopsis
This paper describes the use of a novel porous matrix, Porocell, for high density, tissue-like culture of two insulinoma cell lines, CRI-D2 and CRI-D11. Both these cell lines have previously been shown not to secrete insulin in response to glucose. Porocell is a macro-porous, polymeric material manufactured in the shape of discs that are 6É2 mm in diameter and 2 mm in thickness. Insulinoma cells were cultured in two di †erent mini-bioreactors, each containing six Porocell discs inoculated with 2É5 ] 106 cells per disc. In surface aerated, stirred bioreactors, the insulinoma cells grew as closely packed dense cell sheets penetrating deep into the pores of Porocell. In a second type of system, a packedbed perfused mini-bioreactor, Ñat, extended monolayers of cells were observed growing throughout the Porocell matrix. In both bioreactor conÐgurations, viable cell populations were maintained for 30 days because of the excellent oxygen and nutrient transfer properties of Porocell. CRI-D2 insulinoma cells cultured in static Ñasks and on Porocell did not show any insulin secretion in response to 30 min exposures in media supplemented with 5É5È16É7 mmol dm~3 glucose. However, in long term (14È19 day) cultures, CRI-D2 cells growing in Porocell secreted low, but measurable amounts (25È35 pmol dm~3) of insulin in medium supplemented with elevated (14É5 mmol dm~3) glucose concentrations. The glucose uptake rates of cells cultured in 4É0 mmol dm~3 glucose increased linearly from 1É0 to 2É3 mmol dm~3 day~1 over a period of 19 days. At 14É5 mmol dm~3 glucose concentration, the uptake rate increased from 1É0 to 7É05 mmol dm~3 day~1 over the same period of culture. Contrary to previous studies, we have demonstrated that the CRI-D2 cell line cultured at high cell density in Porocell is capable of secreting insulin when exposed to prolonged and elevated concentrations of glucose. The Porocell mini-bioreactors are easy to use, robust systems that can be used for long-term studies of primary and tumorgenic islet cell function and response to secretagogues.
1998 SCI.