TelematiV1, Ncbi, Checkpoint, bibRecord, 000748

Viability assessment of fresh and frozen/thawed isolated human follicles: reliability of two methods (Trypan blue and Calcein AM/ethidium homodimer-1)

Identifieur interne : 000748 ( Ncbi/Checkpoint ); précédent : 000747; suivant : 000749

Viability assessment of fresh and frozen/thawed isolated human follicles: reliability of two methods (Trypan blue and Calcein AM/ethidium homodimer-1)

Auteurs : Sandra Sanfilippo ; Michel Canis ; Lemlih Ouchchane ; Revaz Botchorishvili ; Christine Artonne ; Laurent Janny ; Florence Brugnon

Source :

Journal of Assisted Reproduction and Genetics [ 1058-0468 ] ; 2011.

RBID : PMC:3241833

Abstract

Purpose

To assess the reliability of trypan blue (TB) and calcein AM/ethidium homodimer-1 (CaAM/EthD-1) staining to evaluate the viability of fresh and thawed human ovarian follicles.

Methods

Isolated follicles from fresh and thawed cortex were stained using TB versus CaAM/EthD-1 methods (n = 10 patients). Measurements were performed by two independent observers. The reliability was evaluated by intraclass correlation coefficient (ICC) and the differences between paired measurements were tested by the Wilcoxon test.

Results

Inter-observer reliability was excellent for each method. Nevertheless, it was even better with the TB method (ICC = 0.83) compared with CaAM/EthD-1 (ICC = 0.75). Moreover, the ICCs for viability measurements using the two methods were good for each observer (observer 1: ICC = 0.49; observer 2: ICC = 0.40).

Conclusion

Compared with CaAM/EthD-1, TB appears to be more reliable as a staining method for follicle viability evaluation. TB staining is a quick and useful method, complementary to histological analysis for quality control in ovarian tissue cryopreservation.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3241833

DOI: 10.1007/s10815-011-9649-y
PubMed: 22033548
PubMed Central: 3241833

Affiliations:

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PMC:3241833

Le document en format XML

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<wicri:noCountry code="subfield">Cedex 1 France</wicri:noCountry>
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<front><div type="abstract" xml:lang="en"><sec><title>Purpose</title>
<p>To assess the reliability of trypan blue (TB) and calcein AM/ethidium homodimer-1 (CaAM/EthD-1) staining to evaluate the viability of fresh and thawed human ovarian follicles.</p>
</sec>
<sec><title>Methods</title>
<p>Isolated follicles from fresh and thawed cortex were stained using TB <italic>versus</italic>
 CaAM/EthD-1 methods (<italic>n</italic>
 = 10 patients). Measurements were performed by two independent observers. The reliability was evaluated by intraclass correlation coefficient (ICC) and the differences between paired measurements were tested by the Wilcoxon test.</p>
</sec>
<sec><title>Results</title>
<p>Inter-observer reliability was excellent for each method. Nevertheless, it was even better with the TB method (ICC = 0.83) compared with CaAM/EthD-1 (ICC = 0.75). Moreover, the ICCs for viability measurements using the two methods were good for each observer (observer 1: ICC = 0.49; observer 2: ICC = 0.40).</p>
</sec>
<sec><title>Conclusion</title>
<p>Compared with CaAM/EthD-1, TB appears to be more reliable as a staining method for follicle viability evaluation. TB staining is a quick and useful method, complementary to histological analysis for quality control in ovarian tissue cryopreservation.</p>
</sec>
</div>
</front>
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<affiliations><list></list>
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<name sortKey="Brugnon, Florence" sort="Brugnon, Florence" uniqKey="Brugnon F" first="Florence" last="Brugnon">Florence Brugnon</name>
<name sortKey="Canis, Michel" sort="Canis, Michel" uniqKey="Canis M" first="Michel" last="Canis">Michel Canis</name>
<name sortKey="Janny, Laurent" sort="Janny, Laurent" uniqKey="Janny L" first="Laurent" last="Janny">Laurent Janny</name>
<name sortKey="Ouchchane, Lemlih" sort="Ouchchane, Lemlih" uniqKey="Ouchchane L" first="Lemlih" last="Ouchchane">Lemlih Ouchchane</name>
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Viability assessment of fresh and frozen/thawed isolated human follicles: reliability of two methods (Trypan blue and Calcein AM/ethidium homodimer-1)

Viability assessment of fresh and frozen/thawed isolated human follicles: reliability of two methods (Trypan blue and Calcein AM/ethidium homodimer-1)

Source :

Abstract

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Links to Exploration step

Le document en format XML

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Pour générer des pages wiki