Fluorescein labelled annexin v staining limitations for determination of blastodermal cells apoptosis

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62156489%3A43210%2F22%3A43920897" target="_blank" >RIV/62156489:43210/22:43920897 - isvavai.cz</a>

Result on the web

<a href="https://doi.org/10.15414/jmbfs.5153" target="_blank" >https://doi.org/10.15414/jmbfs.5153</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.55251/jmbfs.5153" target="_blank" >10.55251/jmbfs.5153</a>

Result language

angličtina

Original language name

Fluorescein labelled annexin v staining limitations for determination of blastodermal cells apoptosis

Original language description

It is known that blastoderm contains not only blastodermal cells (BCs) but also cellular &quot;debris&quot;. These particles include yolk granules and lipid droplets. Some authors used AnnexinV (AnV) staining technique for determination of cell apoptosis. However, AnV is a protein with high affinity to lipids. So the aim of our study was to demonstrate the AnV binding in the blastoderm and yolk cell suspensions. Cell populations were obtained from fertilized chicken eggs. The blastoderms were isolated using filter paper ring method and mechanically fragmented. After that, samples were stained with AnV-FITC and analyzed by fluorescent microscopy (FM) and flow cytometry (FC). FM showed non-specific binding of AnV-FITC on the yolk and lipid particles. Therefore, the FC method using specific nuclear dye DRAQ5 was used to separate BCs from cellular debris. Only 1.82 +/- 0.27 % cells were DRAQ5 positive. On the other hand, increase of total events (21 +/- 3.30 %) of AnV positive cells without DRAQ5 dye was recorded. Moreover, the affinity of yolk to AnV by FC was also observed (10.45 +/- 2.59 %). Significant differences among the groups (P &lt; 0.001) were recorded. This trend was caused by non-specific binding of AnV to the lipid and yolk particles. So it is necessary to stain suspension with nuclear dye to separate BCs than co-stained with some marker of apoptosis. Based on the results we can summarized that AnV staining is inappropriate technique for apoptosis of BCs, however, using nuclear dye we can prevent of non-specific binding.

Czech name

—

Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10601 - Cell biology

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2022

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Microbiology, Biotechnology and Food Sciences

ISSN

1338-5178

e-ISSN

1338-5178

Volume of the periodical

11

Issue of the periodical within the volume

5

Country of publishing house

SK - SLOVAKIA

Number of pages

5

Pages from-to

"e5153"

UT code for WoS article

000719760500001

EID of the result in the Scopus database

2-s2.0-85129245272