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 Citation (Scopus)

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

Fingerprint

Density Gradient Centrifugation
Centrifugation
Mitochondria
insulin
Insulin
lysosomes
Lysosomes
Membrane structures
mitochondria
cells
extracellular space
secretory granules
Secretory Vesicles
Extracellular Space
centrifugation
Adsorption
adsorption
Enzymes
Membranes
Percoll

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

Cite this

Isolation of a purified mitochondrial fraction from viable clonal insulin-producing cells (RINm5F) by percoll density gradient centrifugation. / Abrahamsson, Håkan; Anderson, Tommy; Berggren, Per Olof; Pertoft, Håkan.

In: In Vitro Cellular & Developmental Biology, Vol. 21, No. 7, 01.07.1985, p. 415-420.

Research output: Contribution to journalArticle

Abrahamsson, Håkan ; Anderson, Tommy ; Berggren, Per Olof ; Pertoft, Håkan. / Isolation of a purified mitochondrial fraction from viable clonal insulin-producing cells (RINm5F) by percoll density gradient centrifugation. In: In Vitro Cellular & Developmental Biology. 1985 ; Vol. 21, No. 7. pp. 415-420.
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