TY - JOUR
T1 - Covalent immobilization of luminescent oxygen indicators reduces cytotoxicity
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 -