Cryopreservation of Multicellular Spheroids Derived from Primary Culture of Newborn Piglet Spinal Ganglion Cells
Dorsal root ganglia (DRG) are a potential source of neural stem cells, since they contain the neural crest-derived cells capable to diﬀerentiate into neurons and diﬀerent subpopulations of glial cells. It is known that under certain culture conditions, the DRG cells, derived from neonatal piglets form the ﬂoating multicellular spheroids (MSs), capable to produce the ﬁbroblast-like cells (FLCs), glial cells (GCs) and neuron-like cells (NLCs). Here, we have cryopreserved the MSc, derived under serum-free culture conditions using 5, 7.5, 10% dimethyl sulfoxide (DMSO) and two-stage regimen of freezing. After cryopreservation, the MSs were established to preserve the capability to adhere as well as to produce three cell types: FLCs, GCs and NLCs. However, the capability of frozen-thawed MSs to produce diﬀerent cell types was reduced. The cryoprotectant concentration aﬀected the cell type, prevailing in the culture, derived from the frozen-thawed MSs. After cryopresevation with 10% DMSO, the monolayer consisted mainly of FLCs, while with 5 and 7.5% DMSO it mostly comprised GLCs.
Probl Cryobiol Cryomed 2019; 29(4): 354-364
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