In vitro culture of leukemic cells in collagen scaffolds and
carboxymethyl cellulose-polyethylene glycol gel

J 2024

In vitro culture of leukemic cells in collagen scaffolds and carboxymethyl cellulose-polyethylene glycol gel

SVOZILOVÁ, Hana, Lucy VOJTOVA, Jana MATULOVA, Jana BRUKNEROVÁ, Veronika POLAKOVA et. al.

Basic information

Original name

In vitro culture of leukemic cells in collagen scaffolds and carboxymethyl cellulose-polyethylene glycol gel

Authors

SVOZILOVÁ, Hana (203 Czech Republic, belonging to the institution), Lucy VOJTOVA (203 Czech Republic), Jana MATULOVA (203 Czech Republic), Jana BRUKNEROVÁ (203 Czech Republic, belonging to the institution), Veronika POLAKOVA (203 Czech Republic), Lenka RADOVÁ (203 Czech Republic, belonging to the institution), Michael DOUBEK (203 Czech Republic, belonging to the institution), Karla PLEVOVÁ (203 Czech Republic, belonging to the institution) and Šárka POSPÍŠILOVÁ (203 Czech Republic, belonging to the institution)

Edition

PeerJ, LONDON, PEERJ INC, 2024, 2167-8359

Other information

Language

English

Type of outcome

Article in a journal

Field of Study

30205 Hematology

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

is not subject to a state or trade secret

References:

URL

Impact factor

Impact factor: 2.300 in 2023

RIV identification code

RIV/00216224:14110/24:00138795

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.7717/peerj.18637

UT WoS

001374749200001

Keywords in English

Chronic lymphocytic leukemia; CLL; 3D culture; Carboxymethyl cellulose; CMC; Polyethylene glycol; PEG; Collagen; Scaffolds; Gel

Abstract

V originále

Background: Chronic lymphocytic leukemia (CLL) is a common adult leukemia characterized by the accumulation of neoplastic mature B cells in blood, bone marrow, lymph nodes, and spleen. The disease biology remains unresolved in many aspects, including the processes underlying the disease progression and relapses. However, studying CLL in vitro poses a considerable challenge due to its complexity and dependency on the microenvironment. Several approaches are utilized to overcome this issue, such as co-culture of CLL cells with other cell types, supplementing culture media with growth factors, or setting up a three-dimensional (3D) culture. Previous studies have shown that 3D cultures, compared to conventional ones, can lead to enhanced cell survival and altered gene expression. 3D cultures can also give valuable information while testing treatment response in vitro since they mimic the cell spatial organization more accurately than conventional culture. Methods: In our study, we investigated the behavior of CLL cells in two types of material: (i) solid porous collagen scaffolds and (ii) gel composed of carboxymethyl cellulose and polyethylene glycol (CMC-PEG). We studied CLL cells' distribution, morphology, and viability in these materials by a transmitted-light and confocal microscopy. We also measured the metabolic activity of cultured cells. Additionally, the expression levels of MYC, VCAM1, MCL1, CXCR4, and CCL4 genes in CLL cells were studied by qPCR to observe whether our novel culture approaches lead to increased adhesion, lower apoptotic rates, or activation of cell signaling in relation to the enhanced contact with co-cultured cells. Results: Both materials were biocompatible, translucent, and permeable, as assessed by metabolic assays, cell staining, and microscopy. While collagen scaffolds featured easy manipulation, washability, transferability, and biodegradability, CMC-PEG was advantageous for its easy preparation process and low variability in the number of accommodated cells. Both materials promoted cell-to-cell and cell-to-matrix interactions due to the scaffold structure and generation of cell aggregates. The metabolic activity of CLL cells cultured in CMC-PEG gel was similar to or higher than in conventional culture. Compared to the conventional culture, there was (i) a lower expression of VCAM1 in both materials, (ii) a higher expression of CCL4 in collagen scaffolds, and (iii) a lower expression of CXCR4 and MCL1 (transcript variant 2) in collagen scaffolds, while it was higher in a CMC-PEG gel. Hence, culture in the material can suppress the expression of a pro-apoptotic gene ( MCL1 in collagen scaffolds) or replicate certain gene expression patterns attributed to CLL cells in lymphoid organs (low CXCR4, high CCL4 in collagen scaffolds) or blood (high CXCR4 in CMC-PEG).

Links

LX22NPO5102, research and development project

Name: Národní ústav pro výzkum rakoviny (Acronym: NÚVR)

Investor: Ministry of Education, Youth and Sports of the CR, National institute for cancer research, 5.1 EXCELES

90250, large research infrastructures

Name: Czech-BioImaging III

90251, large research infrastructures

Name: CzechNanoLab II

90253, large research infrastructures

Name: EATRIS-CZ IV

90267, large research infrastructures

Name: NCMG III

Files attached

In_vitro_culture_of_leukemic_cells_in_collagen_scaffolds_and_carboxymethyl_cellulose-polyethylene_glycol_gel.pdf

Cite

SVOZILOVÁ, Hana, Lucy VOJTOVA, Jana MATULOVA, Jana BRUKNEROVÁ, Veronika POLAKOVA, Lenka RADOVÁ, Michael DOUBEK, Karla PLEVOVÁ and Šárka POSPÍŠILOVÁ. In vitro culture of leukemic cells in collagen scaffolds and carboxymethyl cellulose-polyethylene glycol gel. PeerJ. LONDON: PEERJ INC, 2024, vol. 12, December 2024, p. 1-30. ISSN 2167-8359. Available from: https://dx.doi.org/10.7717/peerj.18637.

@article{2475058,
   author = {Svozilová, Hana and Vojtova, Lucy and Matulova, Jana and Bruknerová, Jana and Polakova, Veronika and Radová, Lenka and Doubek, Michael and Plevová, Karla and Pospíšilová, Šárka},
   article_location = {LONDON},
   article_number = {December 2024},
   doi = {http://dx.doi.org/10.7717/peerj.18637},
   keywords = {Chronic lymphocytic leukemia; CLL; 3D culture; Carboxymethyl cellulose; CMC; Polyethylene glycol; PEG; Collagen; Scaffolds; Gel},
   language = {eng},
   issn = {2167-8359},
   journal = {PeerJ},
   title = {In vitro culture of leukemic cells in collagen scaffolds and carboxymethyl cellulose-polyethylene glycol gel},
   url = {https://peerj.com/articles/18637/},
   volume = {12},
   year = {2024}
}

TY  - JOUR
ID  - 2475058
AU  - Svozilová, Hana - Vojtova, Lucy - Matulova, Jana - Bruknerová, Jana - Polakova, Veronika - Radová, Lenka - Doubek, Michael - Plevová, Karla - Pospíšilová, Šárka
PY  - 2024
TI  - In vitro culture of leukemic cells in collagen scaffolds and carboxymethyl cellulose-polyethylene glycol gel
JF  - PeerJ
VL  - 12
IS  - December 2024
SP  - 1-30
EP  - 1-30
PB  - PEERJ INC
SN  - 21678359
KW  - Chronic lymphocytic leukemia
KW  - CLL
KW  - 3D culture
KW  - Carboxymethyl cellulose
KW  - CMC
KW  - Polyethylene glycol
KW  - PEG
KW  - Collagen
KW  - Scaffolds
KW  - Gel
UR  - https://peerj.com/articles/18637/
N2  - Background: Chronic lymphocytic leukemia (CLL) is a common adult leukemia characterized by the accumulation of neoplastic mature B cells in blood, bone marrow, lymph nodes, and spleen. The disease biology remains unresolved in many aspects, including the processes underlying the disease progression and relapses. However, studying CLL in vitro poses a considerable challenge due to its complexity and dependency on the microenvironment. Several approaches are utilized to overcome this issue, such as co-culture of CLL cells with other cell types, supplementing culture media with growth factors, or setting up a three-dimensional (3D) culture. Previous studies have shown that 3D cultures, compared to conventional ones, can lead to enhanced cell survival and altered gene expression. 3D cultures can also give valuable information while testing treatment response in vitro since they mimic the cell spatial organization more accurately than conventional culture. Methods: In our study, we investigated the behavior of CLL cells in two types of material: (i) solid porous collagen scaffolds and (ii) gel composed of carboxymethyl cellulose and polyethylene glycol (CMC-PEG). We studied CLL cells' distribution, morphology, and viability in these materials by a transmitted-light and confocal microscopy. We also measured the metabolic activity of cultured cells. Additionally, the expression levels of MYC, VCAM1, MCL1, CXCR4, and CCL4 genes in CLL cells were studied by qPCR to observe whether our novel culture approaches lead to increased adhesion, lower apoptotic rates, or activation of cell signaling in relation to the enhanced contact with co-cultured cells. Results: Both materials were biocompatible, translucent, and permeable, as assessed by metabolic assays, cell staining, and microscopy. While collagen scaffolds featured easy manipulation, washability, transferability, and biodegradability, CMC-PEG was advantageous for its easy preparation process and low variability in the number of accommodated cells. Both materials promoted cell-to-cell and cell-to-matrix interactions due to the scaffold structure and generation of cell aggregates. The metabolic activity of CLL cells cultured in CMC-PEG gel was similar to or higher than in conventional culture. Compared to the conventional culture, there was (i) a lower expression of VCAM1 in both materials, (ii) a higher expression of CCL4 in collagen scaffolds, and (iii) a lower expression of CXCR4 and MCL1 (transcript variant 2) in collagen scaffolds, while it was higher in a CMC-PEG gel. Hence, culture in the material can suppress the expression of a pro-apoptotic gene ( MCL1 in collagen scaffolds) or replicate certain gene expression patterns attributed to CLL cells in lymphoid organs (low CXCR4, high CCL4 in collagen scaffolds) or blood (high CXCR4 in CMC-PEG).
ER  -

SVOZILOVÁ, Hana, Lucy VOJTOVA, Jana MATULOVA, Jana BRUKNEROVÁ, Veronika POLAKOVA, Lenka RADOVÁ, Michael DOUBEK, Karla PLEVOVÁ and Šárka POSPÍŠILOVÁ. In vitro culture of leukemic cells in collagen scaffolds and carboxymethyl cellulose-polyethylene glycol gel. \textit{PeerJ}. LONDON: PEERJ INC, 2024, vol.~12, December 2024, p.~1-30. ISSN~2167-8359. Available from: https://dx.doi.org/10.7717/peerj.18637.

Přehled o publikaci