Two complementary approaches for efficient isolation of Sertoli cells for transcriptomic analysis
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68378050%3A_____%2F22%3A00566161" target="_blank" >RIV/68378050:_____/22:00566161 - isvavai.cz</a>
Result on the web
<a href="https://www.frontiersin.org/articles/10.3389/fcell.2022.972017/full" target="_blank" >https://www.frontiersin.org/articles/10.3389/fcell.2022.972017/full</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.3389/fcell.2022.972017" target="_blank" >10.3389/fcell.2022.972017</a>
Alternative languages
Result language
angličtina
Original language name
Two complementary approaches for efficient isolation of Sertoli cells for transcriptomic analysis
Original language description
Sertoli cells (SCs) are the only somatic cells that reside in seminiferous tubules of testis. They directly interact with and support the development of germ cells, thus have an indispensable role in the process of spermatogenesis. SCs first appear in a proliferative state and then, with the initiation of the first wave of spermatogenesis, progress to a mature ´nurturing ´ state which supports lifelong continuous sperm production. During this development, the SC transcriptome must adapt rapidly as obstacles in SC maturation often result in deficiencies in male fertility. Due to its importance in spermatogenesis, a reliable, rapid, and precise method for the isolation of high purity, viable and unadulterated SC has been largely missing. We have developed an improved method for the preparation of a testicular single cell suspension comprised of two alternative protocols to separate SCs from the rest of the testicular cells by FACS. The first sorting scheme is based on their co-expression of surface specific markers, FSHr and Occludin-1, while the second focuses on the co-staining of SCs with FSHr-specific antibody and Hoechst 33342, which discriminates DNA content of testicular cells. The entire procedure can be completed in less than 3 h which permits the analysis of the development-related transcriptional profile of these cells. Notably, our comparative study showed that this method resulted in a SC transcriptome that is largely comparable to SCs which were briskly isolated due to their cell-specific expression of fluorescent protein. Interestingly, we also show that SCs sorted as FSHr(+)Occludin(+) cells contained a tangible portion of transcripts from all types of testicular germ cells. Sorting of SCs according to their 2C DNA content significantly reduced the presence of these transcripts, thus seems to be the most suitable approach for accurate determination of the SC transcriptome. We believe that these novel approaches for the isolation of SCs will assist researchers in the elucidation of their function as well as their role in spermatogenesis and disorders related to male infertility.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10601 - Cell biology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2022
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Frontiers in Cell and Developmental Biology
ISSN
2296-634X
e-ISSN
2296-634X
Volume of the periodical
10
Issue of the periodical within the volume
September
Country of publishing house
CH - SWITZERLAND
Number of pages
12
Pages from-to
972017
UT code for WoS article
000875044400001
EID of the result in the Scopus database
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