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001-es BibID:BIBFORM091018
Első szerző:Tajti Gábor (gyógyszerész, biofizikus, sejtbiológus)
Cím:Immunomagnetic separation is a suitable method for electrophysiology and ion channel pharmacology studies on T cells / Gabor Tajti, Tibor Gabor Szanto, Agota Csoti, Greta Racz, César Evaristo, Peter Hajdu, Gyorgy Panyi
Dátum:2021
ISSN:1933-6950 1933-6969
Megjegyzések:Ion channels play pivotal role in the physiological and pathological function of immune cells. As immune cells represent a functionally diverse population, subtype-specific functional studies, such as single-cell electrophysiology require proper subset identification and separation. Magneticactivated cell sorting (MACS) techniques provide an alternative to fluorescence-activated cell sorting (FACS), however, the potential impact of MACS-related beads on the biophysical and pharmacological properties of the ion channels were not studied yet. We studied the aforementioned properties of the voltage-gated Kv1.3 K+ channel in activated CD4+ T-cells as well as the membrane capacitance using whole-cell patch-clamp following immunomagnetic positive separation, using the REAlease? kit. This kit allows three experimental configurations: bead-bound configuration, bead-free configuration following the removal of magnetic beads, and the labelfree configuration following removal of CD4 recognizing antibody fragments. As controls, we used FACS separation as well as immunomagnetic negative selection. The membrane capacitance and of the biophysical parameters of Kv1.3 gating, voltage-dependence of steady-state activation and inactivation kinetics of the current were not affected by the presence of MACS-related compounds on the cell surface. We found subtle differences in the activation kinetics of the Kv1.3 current that could not be explained by the presence of MACS-related compounds. Neither the equilibrium block of Kv1.3 by TEA+ or charybdotoxin (ChTx) nor the kinetics of ChTx block are affected by the presence of the magnetics beads on the cell surface. Based on our results MACS is a suitable method to separate cells for studying ion channels in non-excitable cells, such as T-lymphocytes.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Kv1.3
cd4+ T-cell
magneticactivated cell sorting
macs
immunomagnetic separation
fluorescenceactivated cell sorting
facs
Megjelenés:Channels. - 15 : 1 (2021), p. 53-66. -
További szerzők:Szántó Gábor Tibor (1980-) (vegyész) Csóti Ágota (1989-) (biológus) Rácz Gréta Evaristo, César Hajdu Péter (1975-) (biofizikus) Panyi György (1966-) (biofizikus)
Pályázati támogatás:OTKA K119417
Egyéb
EFOP-3.6.2-16-2017-00006
EFOP
GINOP-2.3.2-15-2016-00015
GINOP
NKFIH K128525
Egyéb
Bolyai Research Fellowship
Egyéb
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