Cold storage of biopsies from wild endangered native Chilean species in field conditions and subsequent isolation of primary culture cell lines
- 2 July 2008
- journal article
- research article
- Published by Springer Science and Business Media LLC in In Vitro Cellular & Developmental Biology – Animal
- Vol. 44 (8-9), 309-320
- https://doi.org/10.1007/s11626-008-9124-y
Abstract
We evaluated the possibility of deriving primary cell cultures from tissue biopsies taken in field conditions from six threaten endemic Chilean species of free-ranging mammals. Biopsies were taken either by ear punching or darts fired to animals and hold in hypothermic conditions (4° C) in defined salt solution for time periods ranging from 0 to 7 d before biopsy samples reached the cell culture laboratory. Previously, holding times were evaluated in experimental cows in controlled conditions. Two enzymatic treatments, collagenase alone or collagenase followed by tripsin, were used to disaggregate tissues for cell culture. We found that ear notches and dart-derived biopsies can be storaged at 4° C for 1 wk and still yield primary cultures. For dart-derived biopsies, there was an invert correlation between length of cold storage and cellular viability in culture. Healthy fibroblast cell lines were obtained in 92% of the biopsies taken despite the origin (punch or biopsy). We are not aware of similar study for free-ranging animals, especially for the use of darting system to biopsy wild terrestrial mammals, we believe that our results could help for a more widespread implementation of these procedures in the practice of ex situ conservationKeywords
This publication has 53 references indexed in Scilit:
- HAIR SAMPLING AND GENOTYPING FROM HAIR FOLLICLES: A MINIMALLY-INVASIVE ALTERNATIVE FOR GENETICS STUDIES IN SMALL, MOBILE PINNIPEDS AND OTHER MAMMALSMarine Mammal Science, 2007
- Maintenance of integrity and function of isolated hepatocytes during extended suspension culture at 25°CLiver International, 2003
- Preservation of the human pancreas before islet isolation using a two-layer (UW solution???perfluorochemical) cold storage method1Transplantation, 2002
- Mammalian Cell Injury Induced by Hypothermia the Emerging Role for Reactive Oxygen SpeciesBiological Chemistry, 2002
- Mitochondrial DNA variation, phylogeography and population structure of the Asian elephantHeredity, 2000
- Conservation status, rarity, and geographic priorities for conservation of Chilean mammals: an assessmentBiological Conservation, 1999
- The potential use of “cloning” in the conservation effortZoo Biology, 1997
- A CETACEAN BIOPSY SYSTEM USING LIGHTWEIGHT PNEUMATIC DARTS, AND ITS EFFECT ON THE BEHAVIOR OF KILLER WHALESMarine Mammal Science, 1996
- A METHOD TO COLLECT AND PROCESS SKIN BIOPSIES FOR CELL CULTURE FROM FREE‐RANGING GRAY WHALES (ESCHRICHTIUS ROBUSTUS)Marine Mammal Science, 1988
- A Biopsy System for Large Whales and Its Use for CytogeneticsJournal of Mammalogy, 1987