Technique for cryopreservation of intestinal smooth muscle cells

S. Batista Lobo, M. Denyer, R. C. Gopalan, F. A. Javid

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Attempts were made to develop a simplified procedure for long-term cryopreservation of intestinal smooth muscle cells (ISMC). ISMC were collected from the ileum of Sprague-Dawley neonatal rats through cellular dissociation in trypsin. Cryopreservation method comprised of a rapid 1-step (protocol 1) and a slow 3-step (protocol 2) freezing of ISMC for 1 week. Preparations were thawed and single ISMC were assessed via the comet assay and damaged DNA was quantified through comet tail moment. The control unfrozen ISMC exhibited DNA damage of 2.34 ± 0.35 compared to ISMC cooled via protocol 2 (2.62 ± 0.36) and protocol 1 (10.15 ± 0.72). Thereafter, protocol 2 freezing method was adopted and ISMC were cryopreserved for 1-week, 1-month, and 4-months to analyse the temporal and long-term cryopreservation of ISMC. This revealed a DNA damage of 2.62 ± 0.36 (1-week), 3.81 ± 0.72 (1-month), and 5.1 ± 0.9 (4-months). Gradual cooling is suitable for continuing storage of ISMC and although fluctuation in cryoinjury is observed with time this is considered to reflect cell-to-cell variability.

Original languageEnglish
Pages (from-to)186-189
Number of pages4
JournalCryobiology
Volume57
Issue number2
DOIs
Publication statusPublished - 1 Oct 2008
Externally publishedYes

Fingerprint

Cryopreservation
smooth muscle
cryopreservation
myocytes
Smooth Muscle Myocytes
Muscle
Cells
methodology
Freezing
DNA damage
DNA Damage
DNA
freezing
Comet Assay
Ileum
ileum
rapid methods
Trypsin
trypsin
Sprague Dawley Rats

Cite this

Batista Lobo, S. ; Denyer, M. ; Gopalan, R. C. ; Javid, F. A. / Technique for cryopreservation of intestinal smooth muscle cells. In: Cryobiology. 2008 ; Vol. 57, No. 2. pp. 186-189.
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Technique for cryopreservation of intestinal smooth muscle cells. / Batista Lobo, S.; Denyer, M.; Gopalan, R. C.; Javid, F. A.

In: Cryobiology, Vol. 57, No. 2, 01.10.2008, p. 186-189.

Research output: Contribution to journalArticle

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