title: Neural differentiation potential of sympathoadrenal progenitors derived from fresh and cryopreserved neonatal porcine adrenal glands.
abstract: Stem/progenitor cells are thought to have the potential in the treatment of severe neurodegenerative diseases. Recently, sympathoadrenal progenitors expressing specific markers of neural crest derivatives and capable to differentiate into neurons were discovered in adult bovine and human adrenal glands, but there was no reported data on cryopreservation of sympathoadrenal progenitors. The aim of the present study was to examine the neural differentiation potential of sympathoadrenal progenitors derived from fresh and cryopreserved neonatal porcine adrenal glands. Considering impact of various initial state of frozen biomaterial on cell recovery, we carried out a comparative estimation of cryopreservation outcome both for adrenal tissue fragments and isolated primary cells. The estimation consisted of determining cell yield, viability, ability to adhere, proliferate and differentiate in vitro. Cells isolated from the fresh adrenal glands were cultured until confluence. A formation of sympathoadrenal progenitors-embedded spherical cell colonies, whose cells are differentiated then into βIII-tubulin-positive cells with neuron-like morphology, was observed on the monolayer. The colonies were well preserved after cryopreservation of cell culture with a cooling rate of 1 °C/min in the cryoprotectant media containing 5-15% of dimethylsulfoxide. Adrenal tissue fragments were cryopreserved in the presence of 10% dimethylsulfoxide at the cooling rates of 0.3; 1: 5; 40 and > 100 °C/min. Sympathoadrenal progenitors were recovered after cryopreservation with 0.3 °C/min cooling rate but not higher.
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine. Electronic address: bozhokgaru@mail.ru.
authors.forename: G A
authors.initials: GA
authors.lastname: Bozhok
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine.
authors.forename: O S
authors.initials: OS
authors.lastname: Sidorenko
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine.
authors.forename: E M
authors.initials: EM
authors.lastname: Plaksina
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine.
authors.forename: T M
authors.initials: TM
authors.lastname: Gurina
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine.
authors.forename: A N
authors.initials: AN
authors.lastname: Sukach
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine.
authors.forename: V S
authors.initials: VS
authors.lastname: Kholodnyy
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine.
authors.forename: V D
authors.initials: VD
authors.lastname: Ustichenko
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine.
authors.forename: S B
authors.initials: SB
authors.lastname: Bilyavskaya
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine.
authors.forename: T P
authors.initials: TP
authors.lastname: Bondarenko
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine.
authors.forename: E I
authors.initials: EI
authors.lastname: Legach
mesh_terms: D000311:Adrenal Glands / Q000166:cytology*; D000818:Animals; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D009432:Neural Crest / Q000166:cytology; D055495:Neurogenesis / Q000502:physiology*; D009474:Neurons / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*; D013552:Swine; D014404:Tubulin / Q000378:metabolism
chemical_list: D003451:Cryoprotective Agents; D014404:Tubulin; D004121:Dimethyl Sulfoxide
keywords: Adrenal glands; Cryopreservation; Dimethylsulfoxide; Primary culture; Sympathoadrenal progenitors; βIII-tubulin