Cardiomyocyte syncytium  combined with Atomic force 
microscopy, advanced setup  of universal biosensor for 
phenotype screening

a 2016

Cardiomyocyte syncytium combined with Atomic force microscopy, advanced setup of universal biosensor for phenotype screening

PEŠL, Martin, Jan PŘIBYL, Šárka JELÍNKOVÁ, Ivana AĆIMOVIĆ, Anton SALYKIN et. al.

Základní údaje

Originální název

Cardiomyocyte syncytium combined with Atomic force microscopy, advanced setup of universal biosensor for phenotype screening

Autoři

PEŠL, Martin, Jan PŘIBYL, Šárka JELÍNKOVÁ, Ivana AĆIMOVIĆ, Anton SALYKIN, Petr DVOŘÁK, Vladimír ROTREKL a Petr SKLÁDAL

Vydání

AFM BioMed Conference, Porto, 2016

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

Biotechnologie a bionika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Organizační jednotka

Lékařská fakulta

Klíčová slova anglicky

AFM biosensor Human stem cell Cardiomyocyte contraction Drug testing

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 13. 6. 2016 15:05, Ing. Mgr. Věra Pospíšilíková

Anotace

V originále

Cardiomyocyte contraction and relaxation are important parameters of cardiac function altered in most cardiac diseases. However we routinely model such diseases using cardiomyocytes derived from human embryonic stem cells and especially from patient specific induced pluripotent stem cells; it is the biosensing of contraction and relaxation which makes the models a great tool for drug development. We employ atomic force microscopy to analyze these parameters. In this novel approach we employ the direct contact of the cantilever with standardized relaxation and stress conditions with the stem cells derived cardiac syncytium. We present advanced experimental setup for quantification of the cardiomyocyte dynamic changes induced by external stimuli such as extreme temperature, calcium level and stress conditions or administered various drugs. Further we show novel automatized data evaluation process. Advantages of feedback- gain parameter previously described (2) are discussed. Methods: Cardiomyocytes were generated as previously described (1) using aggregation in agarose micro-wells allowing homogeneous cluster formation in size and shape (size of the cluster as well as cardiomyocyte content was quantified). Atomic force microscopy analysis was performed in Tyrode’s solution, in Calcium concentration gradients, temperature gradients and ion channel- and adrenergic receptor-modulator gradients.. For data postprocessing and statistical evaluation was involved specific script allowing automatized evaluation. Results: The active AFM mode has no effect on contraction frequency, while providing for correct contraction force recording. We found the optimal value corresponds to average recorded value of EBs contraction force. It was set to 6 nN, in our case, which allowed for the measurement of 95% of maximum contraction force (4.6 nN vs. 4.9 nN, when SP was 6 and 18 nN, respectively). Use of noncoated AFM probes eliminates thermal instability and allows measurements for hours and days continuously. Calcium gradients allowed us to study the excitation-contraction (un)coupling as well as arrythmogenicity of calcium extreme concentrations. Beat to beat surface mapping has shown comparable values all over accessible syncytium rendering the method robust and unbiased by the cantilever position.

Návaznosti

GA13-19910S, projekt VaV

Název: Studium dilatační kardiomyopatie spojené s Duchenovou svalovou dystrofií na modelové tkáni tvořené kardiomyocyty derivovanými z iPS buněk

Investor: Grantová agentura ČR, Studium dilatační kardiomyopatie spojené s Duchenovou svalovou dystrofií na modelové tkáni tvořené kardiomyocyty derivovanými z iPS buněk

Citovat

PEŠL, Martin, Jan PŘIBYL, Šárka JELÍNKOVÁ, Ivana AĆIMOVIĆ, Anton SALYKIN, Petr DVOŘÁK, Vladimír ROTREKL a Petr SKLÁDAL. Cardiomyocyte syncytium combined with Atomic force microscopy, advanced setup of universal biosensor for phenotype screening. In AFM BioMed Conference, Porto. 2016.

@proceedings{1347026,
   author = {Pešl, Martin and Přibyl, Jan and Jelínková, Šárka and Aćimović, Ivana and Salykin, Anton and Dvořák, Petr and Rotrekl, Vladimír and Skládal, Petr},
   booktitle = {AFM BioMed Conference, Porto},
   keywords = {AFM biosensor Human stem cell Cardiomyocyte contraction Drug testing},
   language = {eng},
   title = {Cardiomyocyte syncytium combined with Atomic force microscopy, advanced setup of universal biosensor for phenotype screening},
   url = {http://www.afmbiomed.org/Data/Sites/1/porto-2016/abstractbook_afmbiomed2016_11.04.2016.pdf},
   year = {2016}
}

TY  - CONF
ID  - 1347026
AU  - Pešl, Martin - Přibyl, Jan - Jelínková, Šárka - Aćimović, Ivana - Salykin, Anton - Dvořák, Petr - Rotrekl, Vladimír - Skládal, Petr
PY  - 2016
TI  - Cardiomyocyte syncytium combined with Atomic force microscopy, advanced setup of universal biosensor for phenotype screening
KW  - AFM biosensor Human stem cell Cardiomyocyte contraction Drug testing
UR  - http://www.afmbiomed.org/Data/Sites/1/porto-2016/abstractbook_afmbiomed2016_11.04.2016.pdf
N2  - Cardiomyocyte contraction and relaxation are important parameters of cardiac function altered in most cardiac diseases. However we routinely model such diseases using cardiomyocytes derived from human embryonic stem cells and especially from patient specific induced pluripotent stem cells; it is the biosensing of contraction and relaxation which makes the models a great tool for drug development. We employ atomic force microscopy to analyze these parameters. In this novel approach we employ the direct contact of the cantilever with standardized relaxation and stress conditions with the stem cells derived cardiac syncytium. We present advanced experimental setup for quantification of the cardiomyocyte dynamic changes induced by external stimuli such as extreme temperature, calcium level and stress conditions or administered various drugs. Further we show novel automatized data evaluation process. Advantages of feedback- gain parameter previously described (2) are discussed. Methods: Cardiomyocytes were generated as previously described (1) using aggregation in agarose micro-wells allowing homogeneous cluster formation in size and shape (size of the cluster as well as cardiomyocyte content was quantified). Atomic force microscopy analysis was performed in Tyrode’s solution, in Calcium concentration gradients, temperature gradients and ion channel- and adrenergic receptor-modulator gradients.. For data postprocessing and statistical evaluation was involved specific script allowing automatized evaluation. Results: The active AFM mode has no effect on contraction frequency, while providing for correct contraction force recording. We found the optimal value corresponds to average recorded value of EBs contraction force. It was set to 6 nN, in our case, which allowed for the measurement of 95% of maximum contraction force (4.6 nN vs. 4.9 nN, when SP was 6 and 18 nN, respectively). Use of noncoated AFM probes eliminates thermal instability and allows measurements for hours and days continuously. Calcium gradients allowed us to study the excitation-contraction (un)coupling as well as arrythmogenicity of calcium extreme concentrations. Beat to beat surface mapping has shown comparable values all over accessible syncytium rendering the method robust and unbiased by the cantilever position.
ER  -

PEŠL, Martin, Jan PŘIBYL, Šárka JELÍNKOVÁ, Ivana A$\backslash$'CIMOVI$\backslash$'C, Anton SALYKIN, Petr DVOŘÁK, Vladimír ROTREKL a Petr SKLÁDAL. Cardiomyocyte syncytium combined with Atomic force microscopy, advanced setup of universal biosensor for phenotype screening. In \textit{AFM BioMed Conference, Porto}. 2016.

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