Isolation of a purified mitochondrial fraction from viable clonal insulin-producing cells (RINm5F) by percoll density gradient centrifugation

Håkan Abrahamsson, Tommy Anderson, Per Olof Berggren, Håkan Pertoft

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

A Percoll density gradient was employed for selecting large numbers of viable insulin-producing RINm5F cells. Homogenates of these cells were then subjected to gradient centrifugation and two clearly visible bands were obtained. The light fraction was essentially composed of mitochondria banded at a density of about 1.06 g/ml. The heavier fraction banded at 1.09 to 1.10 g/ml and contained lysosomes and a small number of secretory granules. The distribution of Percoll particles was restricted to the extracellular space and there was no adsorption to any membrane structures. The distribution pattern of marker enzymes for the mitochondria and lysosomes was similar to that of normal pancreatic β-cells. With the use of a Percoll density gradient it was thus possible to isolate a purified mitochondrial fraction from viable RINm5F cells.

Original languageEnglish (US)
Pages (from-to)415-420
Number of pages6
JournalIn Vitro Cellular & Developmental Biology
Volume21
Issue number7
DOIs
StatePublished - Jul 1 1985

Keywords

  • clonal insulin-producing cells (TINm5F)
  • isolation of intact cells
  • Percoll density gradient
  • purified mitochondrial fraction

ASJC Scopus subject areas

  • Biotechnology
  • Plant Science
  • Cell Biology
  • Clinical Biochemistry
  • Developmental Biology

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