Establishment of oral squamous cell carcinoma cell line and
magnetic bead-based isolation and characterization of its
CD90/CD44 subpopulations

J 2017

Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations

SVOBODOVÁ, Markéta, Martina RAUDENSKÁ, Jaromír GUMULEC, Jan BALVAN, Michaela FOJTŮ et. al.

Základní údaje

Originální název

Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations

Autoři

SVOBODOVÁ, Markéta (203 Česká republika, domácí), Martina RAUDENSKÁ (203 Česká republika, domácí), Jaromír GUMULEC (203 Česká republika, domácí), Jan BALVAN (203 Česká republika, domácí), Michaela FOJTŮ (203 Česká republika, domácí), Monika KRATOCHVÍLOVÁ (203 Česká republika, domácí), Hana POLANSKÁ (203 Česká republika, domácí), Zuzana HORÁKOVÁ (203 Česká republika), Rom KOSTŘICA (203 Česká republika), Petr BABULA (203 Česká republika, domácí), Z. HEGER (203 Česká republika) a Michal MASAŘÍK (203 Česká republika, garant, domácí)

Vydání

Oncotarget, Albany, Impact Journals, 2017, 1949-2553

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30204 Oncology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 5.168 v roce 2016

Kód RIV

RIV/00216224:14110/17:00095052

Organizační jednotka

Lékařská fakulta

DOI

http://dx.doi.org/10.18632/oncotarget.19914

UT WoS

000410291200128

Klíčová slova anglicky

head and neck neoplasms; coculture techniques; cell line; tumor; carcinoma

Štítky

EL OK

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 20. 3. 2018 12:43, Soňa Böhmová

Anotace

V originále

In this study, we describe the establishment of the human papillomavirus 18-positive, stage II, grade 1, T2N0M0 head and neck tumor primary cell line derived from oral squamous cell carcinoma of a non-smoking patient by using two different protocols. Furthermore, a preparation of subpopulations derived from this primary cell line according to the cluster of differentiation molecules CD44/CD90 status using magnetic bead-based separation and their characterization was performed. Impedance-based real-time cell analysis, enzyme-linked immunsorbant assay (ELISA), wound-healing assay, flow-cytometry, gene expression analysis, and MTT assay were used to characterize these four subpopulations (CD44(+)/CD90(-), CD44(-)/CD90(-), CD44(+)/CD90(+), CD44(-)/CD90(-)). We optimised methodics for establishement of primary cell lines derived from oral squamous cell carcinoma tissue samples and subsequent separation of mesenchymal (CD90(+)) and epithelial (CD90(-)) types of tumorous cells. Primary cell line prepared by using trypsin proteolysis was more viable than the one prepared by using collagenase. According to our results, CD90 separation is a necessary step in preparation of permanent tumor-tissue derived cell lines. Based on the wound-healing assay, CD44(+) cells exhibited stronger migratory capacity than CD44(-) subpopulations. CD44(+) subpopulations had also significantly higher expression of BIRC5 and SOX2, lower expression of FLT1 and IL6, and higher levels of basal autophagy compared to CD44(-) subpopulations. Furthermore, co-cultivation experiments revealed that CD44(-)/CD90(+) cells supported growth of epithelial tumor cells (CD44(+)/CD90(-)). On the contrary, factors released by CD44(+)/CD90(+) type of cells seem to have rather inhibiting effect. The most cisplatin-resistant subpopulation with the shortest doubling time was CD44(-)/CD90(+), but this subpopulation had a low migratory capacity.

Návaznosti

GA16-12454S, projekt VaV

Název: Charakterizace a modifikace komplexní odpovědi buněk nádorů hlavy a krku na různá záření - krok kupředu ke kombinované personalizované (radio)terapii

Investor: Grantová agentura ČR, Charakterizace a modifikace komplexní odpovědi buněk nádorů hlavy a krku na různá záření - krok kupředu ke kombinované personalizované (radio)terapii

NV16-29835A, projekt VaV

Název: Molekulárně-genetické markery predikce účinnosti radioterapie u nádorů hlavy a krku

ROZV/24/LF/2016, interní kód MU

Název: LF - Příspěvek IP 2016

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, LF - Příspěvek IP 2016

Citovat

SVOBODOVÁ, Markéta, Martina RAUDENSKÁ, Jaromír GUMULEC, Jan BALVAN, Michaela FOJTŮ, Monika KRATOCHVÍLOVÁ, Hana POLANSKÁ, Zuzana HORÁKOVÁ, Rom KOSTŘICA, Petr BABULA, Z. HEGER a Michal MASAŘÍK. Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations. Oncotarget. Albany: Impact Journals, 2017, roč. 8, č. 39, s. 66254-66269. ISSN 1949-2553. Dostupné z: https://dx.doi.org/10.18632/oncotarget.19914.

@article{1392138,
   author = {Svobodová, Markéta and Raudenská, Martina and Gumulec, Jaromír and Balvan, Jan and Fojtů, Michaela and Kratochvílová, Monika and Polanská, Hana and Horáková, Zuzana and Kostřica, Rom and Babula, Petr and Heger, Z. and Masařík, Michal},
   article_location = {Albany},
   article_number = {39},
   doi = {http://dx.doi.org/10.18632/oncotarget.19914},
   keywords = {head and neck neoplasms; coculture techniques; cell line; tumor; carcinoma},
   language = {eng},
   issn = {1949-2553},
   journal = {Oncotarget},
   title = {Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations},
   volume = {8},
   year = {2017}
}

TY  - JOUR
ID  - 1392138
AU  - Svobodová, Markéta - Raudenská, Martina - Gumulec, Jaromír - Balvan, Jan - Fojtů, Michaela - Kratochvílová, Monika - Polanská, Hana - Horáková, Zuzana - Kostřica, Rom - Babula, Petr - Heger, Z. - Masařík, Michal
PY  - 2017
TI  - Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations
JF  - Oncotarget
VL  - 8
IS  - 39
SP  - 66254-66269
EP  - 66254-66269
PB  - Impact Journals
SN  - 19492553
KW  - head and neck neoplasms
KW  - coculture techniques
KW  - cell line
KW  - tumor
KW  - carcinoma
N2  - In this study, we describe the establishment of the human papillomavirus 18-positive, stage II, grade 1, T2N0M0 head and neck tumor primary cell line derived from oral squamous cell carcinoma of a non-smoking patient by using two different protocols. Furthermore, a preparation of subpopulations derived from this primary cell line according to the cluster of differentiation molecules CD44/CD90 status using magnetic bead-based separation and their characterization was performed. Impedance-based real-time cell analysis, enzyme-linked immunsorbant assay (ELISA), wound-healing assay, flow-cytometry, gene expression analysis, and MTT assay were used to characterize these four subpopulations (CD44(+)/CD90(-), CD44(-)/CD90(-), CD44(+)/CD90(+), CD44(-)/CD90(-)). We optimised methodics for establishement of primary cell lines derived from oral squamous cell carcinoma tissue samples and subsequent separation of mesenchymal (CD90(+)) and epithelial (CD90(-)) types of tumorous cells. Primary cell line prepared by using trypsin proteolysis was more viable than the one prepared by using collagenase. According to our results, CD90 separation is a necessary step in preparation of permanent tumor-tissue derived cell lines. Based on the wound-healing assay, CD44(+) cells exhibited stronger migratory capacity than CD44(-) subpopulations. CD44(+) subpopulations had also significantly higher expression of BIRC5 and SOX2, lower expression of FLT1 and IL6, and higher levels of basal autophagy compared to CD44(-) subpopulations. Furthermore, co-cultivation experiments revealed that CD44(-)/CD90(+) cells supported growth of epithelial tumor cells (CD44(+)/CD90(-)). On the contrary, factors released by CD44(+)/CD90(+) type of cells seem to have rather inhibiting effect. The most cisplatin-resistant subpopulation with the shortest doubling time was CD44(-)/CD90(+), but this subpopulation had a low migratory capacity.
ER  -

SVOBODOVÁ, Markéta, Martina RAUDENSKÁ, Jaromír GUMULEC, Jan BALVAN, Michaela FOJTŮ, Monika KRATOCHVÍLOVÁ, Hana POLANSKÁ, Zuzana HORÁKOVÁ, Rom KOSTŘICA, Petr BABULA, Z. HEGER a Michal MASAŘÍK. Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations. \textit{Oncotarget}. Albany: Impact Journals, 2017, roč.~8, č.~39, s.~66254-66269. ISSN~1949-2553. Dostupné z: https://dx.doi.org/10.18632/oncotarget.19914.

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