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Characterization of bioactive substances MHGF-68 on tumour cell lines with LiveFlow In Vitro Technology

R. Hodoši
R. Hodoši
, 
E. Nováková
E. Nováková
, 
K. Macková
K. Macková
, 
M. Molitorisová
M. Molitorisová
 et 
M. Šupolíková
M. Šupolíková
   | 18 juin 2021
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Article Category: Original Paper
Publié en ligne: 18 juin 2021
Pages: 24 - 29
Reçu: 11 nov. 2020
Accepté: 22 mars 2021
DOI: https://doi.org/10.2478/afpuc-2020-0021
Mots clés
© 2021 Hodoši R. et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

LiveFlow system scheme. Blue and green – LiveBox with samples, parallel circuits of medium, Red – controls, series circuit of medium.
LiveFlow system scheme. Blue and green – LiveBox with samples, parallel circuits of medium, Red – controls, series circuit of medium.

Figure 2

Scheme of dynamic cultivation using LiveFlow (IVTech). LiveBox with adherent cell line on the glass slide inside, reservoir with cultivation medium and LiveFlow ensuring continual flow of the medium.
Scheme of dynamic cultivation using LiveFlow (IVTech). LiveBox with adherent cell line on the glass slide inside, reservoir with cultivation medium and LiveFlow ensuring continual flow of the medium.

Figure 3

Distribution of actin filaments in Hepa1c1c7 cells incubated with MHGF-68 after dynamic cultivation.Hepa1c1c7 cells were incubated with MHGF-68. After 0, 24 and 72 h, the cells were fixed and labeled with Alexa Fluor 555 phalloidin. Nuclei were stained with DAPI. The intracelular distribution of actin filamets (green) and nuclei (blue) were imaged by confocal laser scanning fluorescence microscopy (Leica TCS SP8 AOBS) with HC PL APO CS2 63×/1.40 OIL lens.A Hepa1c1c7 untreated, 0 h cultivation B Hepa1c1c7 untreated, 24 h cultivation C Hepa1c1c7 untreated, 72 h cultivation D Hepa1c1c7 + MHGF-68, 0 h cultivation E Hepa1c1c7 + MHGF-68, 24 h cultivation F Hepa1c1c7 + MHGF-68, 72 h cultivation
Distribution of actin filaments in Hepa1c1c7 cells incubated with MHGF-68 after dynamic cultivation.Hepa1c1c7 cells were incubated with MHGF-68. After 0, 24 and 72 h, the cells were fixed and labeled with Alexa Fluor 555 phalloidin. Nuclei were stained with DAPI. The intracelular distribution of actin filamets (green) and nuclei (blue) were imaged by confocal laser scanning fluorescence microscopy (Leica TCS SP8 AOBS) with HC PL APO CS2 63×/1.40 OIL lens.A Hepa1c1c7 untreated, 0 h cultivation B Hepa1c1c7 untreated, 24 h cultivation C Hepa1c1c7 untreated, 72 h cultivation D Hepa1c1c7 + MHGF-68, 0 h cultivation E Hepa1c1c7 + MHGF-68, 24 h cultivation F Hepa1c1c7 + MHGF-68, 72 h cultivation

Figure 4

Distribution of actin filaments in Hepa1c1c7 cells incubated with MHGF-68 after static cultivation.Hepa1c1c7 cells were incubated with MHGF-68. After 0, 24 and 72 h, the cells were fixed and labeled with Alexa Fluor 555 phalloidin.Nuclei were stained with DAPI. The intracelular distribution of actin filamets (green) and nuclei (blue) were imaged by confocal laser scanning fluorescence microscopy (Leica TCS SP8 AOBS) with HC PL APO CS2 63×/1.40 OIL lens. A Hepa1c1c7 untreated, 0 h cultivation B Hepa1c1c7 untreated, 24 h cultivation C Hepa1c1c7 untreated, 72 h cultivation D Hepa1c1c7 + MHGF-68, 0 h cultivation E Hepa1c1c7 + MHGF-68, 24 h cultivation F Hepa1c1c7 + MHGF-68, 72 h cultivation
Distribution of actin filaments in Hepa1c1c7 cells incubated with MHGF-68 after static cultivation.Hepa1c1c7 cells were incubated with MHGF-68. After 0, 24 and 72 h, the cells were fixed and labeled with Alexa Fluor 555 phalloidin.Nuclei were stained with DAPI. The intracelular distribution of actin filamets (green) and nuclei (blue) were imaged by confocal laser scanning fluorescence microscopy (Leica TCS SP8 AOBS) with HC PL APO CS2 63×/1.40 OIL lens. A Hepa1c1c7 untreated, 0 h cultivation B Hepa1c1c7 untreated, 24 h cultivation C Hepa1c1c7 untreated, 72 h cultivation D Hepa1c1c7 + MHGF-68, 0 h cultivation E Hepa1c1c7 + MHGF-68, 24 h cultivation F Hepa1c1c7 + MHGF-68, 72 h cultivation

Figure 5

Distribution of actin filaments in Hepa1c1c7 cells incubated 72 h with MHGF-68 after static and dynamic cultivation.Hepa1c1c7 cells were incubated with MHGF-68. After 72 h, the cells were fixed and labeled with Alexa Fluor 555 phalloidin. Nuclei were stained with DAPI. The intracelular distribution of actin filamets (green) and nuclei (blue) were imaged by confocal laser scanning fluorescence microscopy (Leica TCS SP8 AOBS) with HC PL APO CS2 63×/1.40 OIL lens.A Hepa1c1c7 untreated, static cultivation B Hepa1c1c7 + MHGF-68, static cultivation C Hepa1c1c7 untreated, dynamic cultivation D Hepa1c1c7 + MHGF-68, dynamic cultivation
Distribution of actin filaments in Hepa1c1c7 cells incubated 72 h with MHGF-68 after static and dynamic cultivation.Hepa1c1c7 cells were incubated with MHGF-68. After 72 h, the cells were fixed and labeled with Alexa Fluor 555 phalloidin. Nuclei were stained with DAPI. The intracelular distribution of actin filamets (green) and nuclei (blue) were imaged by confocal laser scanning fluorescence microscopy (Leica TCS SP8 AOBS) with HC PL APO CS2 63×/1.40 OIL lens.A Hepa1c1c7 untreated, static cultivation B Hepa1c1c7 + MHGF-68, static cultivation C Hepa1c1c7 untreated, dynamic cultivation D Hepa1c1c7 + MHGF-68, dynamic cultivation

Comparison of IVTech, in vitro and in vivo testing.

In vitro In vivo IVTech (advanced cell culture systems)
Lack of human complexity Ethically controversial Human organ environment simulation
Lack of side effects tests Time ineffective Multi-organ models
Lack of geometrical complexity Expensive (2–30 times more than in vitro) 3D and dynamic cell cultures
Cells cultivated in static conditions No high-throughput monitoring Real time monitoring
eISSN:
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