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.

Basic information

Original name

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

Authors

SVOBODOVÁ, Markéta (203 Czech Republic, belonging to the institution), Martina RAUDENSKÁ (203 Czech Republic, belonging to the institution), Jaromír GUMULEC (203 Czech Republic, belonging to the institution), Jan BALVAN (203 Czech Republic, belonging to the institution), Michaela FOJTŮ (203 Czech Republic, belonging to the institution), Monika KRATOCHVÍLOVÁ (203 Czech Republic, belonging to the institution), Hana POLANSKÁ (203 Czech Republic, belonging to the institution), Zuzana HORÁKOVÁ (203 Czech Republic), Rom KOSTŘICA (203 Czech Republic), Petr BABULA (203 Czech Republic, belonging to the institution), Z. HEGER (203 Czech Republic) and Michal MASAŘÍK (203 Czech Republic, guarantor, belonging to the institution)

Edition

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

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30204 Oncology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 5.168 in 2016

RIV identification code

RIV/00216224:14110/17:00095052

Organization unit

Faculty of Medicine

DOI

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

UT WoS

000410291200128

Keywords in English

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

Abstract

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.

Links

GA16-12454S, research and development project

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

Investor: Czech Science Foundation

NV16-29835A, research and development project

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

ROZV/24/LF/2016, interní kód MU

Name: LF - Příspěvek IP 2016

Investor: Ministry of Education, Youth and Sports of the CR

Citovat

SVOBODOVÁ, Markéta, Martina RAUDENSKÁ, Jaromír GUMULEC, Jan BALVAN, Michaela FOJTŮ, Monika KRATOCHVÍLOVÁ, Hana POLANSKÁ, Zuzana HORÁKOVÁ, Rom KOSTŘICA, Petr BABULA, Z. HEGER and 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, vol. 8, No 39, p. 66254-66269. ISSN 1949-2553. Available from: 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 and 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, vol.~8, No~39, p.~66254-66269. ISSN~1949-2553. Available from: https://dx.doi.org/10.18632/oncotarget.19914.

Detailed Information on Publication Record