Covalent immobilization of luminescent oxygen indicators reduces cytotoxicity

Hannu Välimäki; Tanja Hyvärinen; Joni Leivo; Haider Iftikhar; Mari Pekkanen-Mattila; Dhanesh Kattipparambil Rajan; Jarmo Verho; Joose Kreutzer; Tomi Ryynänen; Jonatan Pirhonen; Katriina Aalto-Setälä; Pasi Kallio; Susanna Narkilahti; Jukka Lekkala

doi:10.1007/s10544-020-00495-3

Covalent immobilization of luminescent oxygen indicators reduces cytotoxicity

Hannu Välimäki, Tanja Hyvärinen, Joni Leivo, Haider Iftikhar, Mari Pekkanen-Mattila, Dhanesh Kattipparambil Rajan, Jarmo Verho, Joose Kreutzer, Tomi Ryynänen, Jonatan Pirhonen, Katriina Aalto-Setälä, Pasi Kallio, Susanna Narkilahti, Jukka Lekkala

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Abstract

Luminescence-based oxygen sensing is a widely used tool in cell culture applications. In a typical configuration, the luminescent oxygen indicators are embedded in a solid, oxygen-permeable matrix in contact with the culture medium. However, in sensitive cell cultures even minimal leaching of the potentially cytotoxic indicators can become an issue. One way to prevent the leaching is to immobilize the indicators covalently into the supporting matrix. In this paper, we report on a method where platinum(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorphenyl)-porphyrin (PtTFPP) oxygen indicators are covalently immobilized into a polymer matrix consisting of polystyrene and poly(pentafluorostyrene). We study how the covalent immobilization influences the sensing material’s cytotoxicity to human induced pluripotent stem cell-derived (hiPSC-derived) neurons and cardiomyocytes (CMs) through 7–13 days culturing experiments and various viability analyses. Furthermore, we study the effect of the covalent immobilization on the indicator leaching and the oxygen sensing properties of the material. In addition, we demonstrate the use of the covalently linked oxygen sensing material in real time oxygen tension monitoring in functional hypoxia studies of the hiPSC-derived CMs. The results show that the covalently immobilized indicators substantially reduce indicator leaching and the cytotoxicity of the oxygen sensing material, while the influence on the oxygen sensing properties remains small or nonexistent.

Original language	English
Article number	41
Journal	BIOMEDICAL MICRODEVICES
Volume	22
Issue number	2
DOIs	https://doi.org/10.1007/s10544-020-00495-3
Publication status	Published - 1 Jun 2020
Publication type	A1 Journal article-refereed

Keywords

Covalently immobilized indicators
hiPSC-derived neurons and cardiomyocytes
Human induced pluripotent stem cell-derived cells
Luminescent-based oxygen sensing
PtTFPP cytotoxicity

Publication forum classification

Publication forum level 1

ASJC Scopus subject areas

Biomedical Engineering
Molecular Biology

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10.1007/s10544-020-00495-3Licence: CC BY

Välimäki2020_Article_CovalentImmobilizationOfLumineFinal published version, 1.65 MBLicence: CC BY

http://urn.fi/URN:NBN:fi:tuni-202006266236Licence: CC BY

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Välimäki, H., Hyvärinen, T., Leivo, J., Iftikhar, H., Pekkanen-Mattila, M., Rajan, D. K., Verho, J., Kreutzer, J., Ryynänen, T., Pirhonen, J., Aalto-Setälä, K., Kallio, P., Narkilahti, S., & Lekkala, J. (2020). Covalent immobilization of luminescent oxygen indicators reduces cytotoxicity. BIOMEDICAL MICRODEVICES, 22(2), Article 41. https://doi.org/10.1007/s10544-020-00495-3

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title = "Covalent immobilization of luminescent oxygen indicators reduces cytotoxicity",

abstract = "Luminescence-based oxygen sensing is a widely used tool in cell culture applications. In a typical configuration, the luminescent oxygen indicators are embedded in a solid, oxygen-permeable matrix in contact with the culture medium. However, in sensitive cell cultures even minimal leaching of the potentially cytotoxic indicators can become an issue. One way to prevent the leaching is to immobilize the indicators covalently into the supporting matrix. In this paper, we report on a method where platinum(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorphenyl)-porphyrin (PtTFPP) oxygen indicators are covalently immobilized into a polymer matrix consisting of polystyrene and poly(pentafluorostyrene). We study how the covalent immobilization influences the sensing material{\textquoteright}s cytotoxicity to human induced pluripotent stem cell-derived (hiPSC-derived) neurons and cardiomyocytes (CMs) through 7–13 days culturing experiments and various viability analyses. Furthermore, we study the effect of the covalent immobilization on the indicator leaching and the oxygen sensing properties of the material. In addition, we demonstrate the use of the covalently linked oxygen sensing material in real time oxygen tension monitoring in functional hypoxia studies of the hiPSC-derived CMs. The results show that the covalently immobilized indicators substantially reduce indicator leaching and the cytotoxicity of the oxygen sensing material, while the influence on the oxygen sensing properties remains small or nonexistent.",

keywords = "Covalently immobilized indicators, hiPSC-derived neurons and cardiomyocytes, Human induced pluripotent stem cell-derived cells, Luminescent-based oxygen sensing, PtTFPP cytotoxicity",

author = "Hannu V{\"a}lim{\"a}ki and Tanja Hyv{\"a}rinen and Joni Leivo and Haider Iftikhar and Mari Pekkanen-Mattila and Rajan, {Dhanesh Kattipparambil} and Jarmo Verho and Joose Kreutzer and Tomi Ryyn{\"a}nen and Jonatan Pirhonen and Katriina Aalto-Set{\"a}l{\"a} and Pasi Kallio and Susanna Narkilahti and Jukka Lekkala",

year = "2020",

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doi = "10.1007/s10544-020-00495-3",

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AU - Välimäki, Hannu

AU - Hyvärinen, Tanja

AU - Leivo, Joni

AU - Iftikhar, Haider

AU - Pekkanen-Mattila, Mari

AU - Rajan, Dhanesh Kattipparambil

AU - Verho, Jarmo

AU - Kreutzer, Joose

AU - Ryynänen, Tomi

AU - Pirhonen, Jonatan

AU - Aalto-Setälä, Katriina

AU - Kallio, Pasi

AU - Narkilahti, Susanna

AU - Lekkala, Jukka

PY - 2020/6/1

Y1 - 2020/6/1

N2 - Luminescence-based oxygen sensing is a widely used tool in cell culture applications. In a typical configuration, the luminescent oxygen indicators are embedded in a solid, oxygen-permeable matrix in contact with the culture medium. However, in sensitive cell cultures even minimal leaching of the potentially cytotoxic indicators can become an issue. One way to prevent the leaching is to immobilize the indicators covalently into the supporting matrix. In this paper, we report on a method where platinum(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorphenyl)-porphyrin (PtTFPP) oxygen indicators are covalently immobilized into a polymer matrix consisting of polystyrene and poly(pentafluorostyrene). We study how the covalent immobilization influences the sensing material’s cytotoxicity to human induced pluripotent stem cell-derived (hiPSC-derived) neurons and cardiomyocytes (CMs) through 7–13 days culturing experiments and various viability analyses. Furthermore, we study the effect of the covalent immobilization on the indicator leaching and the oxygen sensing properties of the material. In addition, we demonstrate the use of the covalently linked oxygen sensing material in real time oxygen tension monitoring in functional hypoxia studies of the hiPSC-derived CMs. The results show that the covalently immobilized indicators substantially reduce indicator leaching and the cytotoxicity of the oxygen sensing material, while the influence on the oxygen sensing properties remains small or nonexistent.

AB - Luminescence-based oxygen sensing is a widely used tool in cell culture applications. In a typical configuration, the luminescent oxygen indicators are embedded in a solid, oxygen-permeable matrix in contact with the culture medium. However, in sensitive cell cultures even minimal leaching of the potentially cytotoxic indicators can become an issue. One way to prevent the leaching is to immobilize the indicators covalently into the supporting matrix. In this paper, we report on a method where platinum(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorphenyl)-porphyrin (PtTFPP) oxygen indicators are covalently immobilized into a polymer matrix consisting of polystyrene and poly(pentafluorostyrene). We study how the covalent immobilization influences the sensing material’s cytotoxicity to human induced pluripotent stem cell-derived (hiPSC-derived) neurons and cardiomyocytes (CMs) through 7–13 days culturing experiments and various viability analyses. Furthermore, we study the effect of the covalent immobilization on the indicator leaching and the oxygen sensing properties of the material. In addition, we demonstrate the use of the covalently linked oxygen sensing material in real time oxygen tension monitoring in functional hypoxia studies of the hiPSC-derived CMs. The results show that the covalently immobilized indicators substantially reduce indicator leaching and the cytotoxicity of the oxygen sensing material, while the influence on the oxygen sensing properties remains small or nonexistent.

KW - Covalently immobilized indicators

KW - hiPSC-derived neurons and cardiomyocytes

KW - Human induced pluripotent stem cell-derived cells

KW - Luminescent-based oxygen sensing

KW - PtTFPP cytotoxicity

U2 - 10.1007/s10544-020-00495-3

DO - 10.1007/s10544-020-00495-3

M3 - Article

C2 - 32494857

AN - SCOPUS:85085923111

SN - 1387-2176

VL - 22

JO - BIOMEDICAL MICRODEVICES

JF - BIOMEDICAL MICRODEVICES

IS - 2

M1 - 41

ER -

Covalent immobilization of luminescent oxygen indicators reduces cytotoxicity

Abstract

Keywords

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