Isolation of intact nuclei of high purity from mouse liver

Analytical Biochemistry
Takako NagataRaj Lakshman

Abstract

Current methods of nuclear isolation from liver disrupt the plasmalemmae via homogenization and separation of the nuclei by high centrifugal force (HCF) through gradients of sucrose or other substances for up to 80 min. The use of HCF for such a long time increases the potential for nuclear damage and degradation by endogenous proteases. We compared four combinations of alterations to classical nuclear isolation methods as follows. Mouse liver was gently crushed through a fine mesh with and without in vivo perfusion with collagenase. The cell suspension was centrifuged at 600 g to remove gross debris and then at moderate centrifugal force (MCF, 16,000 g) or high centrifugal force (HCF, 70,000 g) through sucrose gradients for 30 min. The purity of the isolated nuclei was assessed biologically and morphologically, including analyses of representative marker proteins for nuclei and cytoplasm. The results indicate that MCF and no collagenase provided the highest nuclear integrity and purity, whereas MCF with collagenase is a viable option if priority is given to yield. The method is especially suited for small samples and so should facilitate studies with human liver biopsies and livers from mice, the most widely used species for g...Continue Reading

References

Jun 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D O Quissell, R S Redman
Jan 1, 1985·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·P M BrannonN Kretchmer
Jan 1, 1985·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·K Bose, D C Allison
Dec 1, 1967·The Journal of Cell Biology·R B HowardL A Pesch
Apr 2, 1998·Journal of Pharmacological and Toxicological Methods·Y F Ho, T M Guenthner
Nov 1, 1960·Stain Technology·K C RICHARDSONE H FINKE
May 1, 1964·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·W C HYMER, E L KUFF

❮ Previous
Next ❯

Citations

Apr 2, 2013·Antioxidants & Redox Signaling·José Luis García-GiménezFederico V Pallardó
Oct 8, 2013·Toxicological Sciences : an Official Journal of the Society of Toxicology·Puttappa R DodmaneSamuel M Cohen

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.

Related Papers

Journal of the Association for Research in Otolaryngology : JARO
Rachel A DumontPeter G Gillespie
Experimental Cell Research
Nathalie Fomproix, Piergiorgio Percipalle
Proceedings of the National Academy of Sciences of the United States of America
A NagyJ C Roder
© 2022 Meta ULC. All rights reserved