Characterization of neuron-like cells derived from canine bone marrow stromal cells

Abstract

Regenerative therapy using bone marrow stromal cells (BMSCs) has begun to be clinically applied in humans and dogs for neurological disorders such as spinal cord injury. Under appropriate conditions in vitro, BMSCs differentiate into neuronal cells, which may improve the effects of regenerative therapy. In this study, we evaluated canine neuron-like cells (NLCs) derived from BMSCs. We speculated on their suitability for neuro-transplantation from the point of view of their morphological features, long-term viability, abundant availability, and ability to be subcultured. Canine NLCs were differentiated as follows: third-passage BMSCs were maintained in pre-induction medium containing 2-mercaptoethanol and dimethylsulfoxide for 5 h, and then cells were transferred to neuronal induction medium containing fetal bovine serum, basic fibroblast growth factor, epidermal growth factor, dibutyryl cyclic AMP, and isobutylmethylxanthine for 7 or 14 days. Canine NLCs fulfilled the transplantation criteria and expressed markers of both immature neurons (nestin, 84.7 %) and mature neuronal cells (microtubule-associated protein-2, 95.7 %; βIII-tubulin protein, 12.9 %; glial fibrillary acidic protein, 9.2 %). These results suggest that canine BMSCs can be induced to differentiate into neuronal cells and may be suitable for neuro-transplantation. This study may provide information for improving cellular therapy for neurological diseases.

Acute Spinal Cord Injury

“…The aim is to restore function by transplanting cells, intrathecally (ie, into the spinal cord) or intravenously, at or near the time of acute SCI, to provide neuroprotection. These data merit attention because obtaining and culturing cells (eg, mesenchymal stem cells [MSCs] from adipose tissue, umbilical cord blood, or bone marrow stromal cells [92][93][94] ) is readily possible nowadays, and it is therefore worth examining how far veterinarians are from translating research protocols from the laboratory to pets in the clinic. Here the data on canine cell transplantation within a month of the trauma is reviewed.…”

Differentiation of canine bone marrow stromal cells into voltage- and glutamate-responsive neuron-like cells by basic fibroblast growth factor

“…In our present study, incubation with bFGF resulted in the maintenance of the viability of canine BMSCs for significant periods in the expression of neuronal markers. Previously, it was reported that many types of mouse, human and canine cells were cultured for long time in the media containing bFGF, indicating that bFGF was involved in the maintenance of viability of the mouse, human and canine cells [1, 13, 24, 33]. Therefore, bFGF might contribute to maintenance of the cell viability during the neuronal differentiation of canine BMSCs.…”

“…In the previous studies on neuronal differentiation of canine BMSCs, it has been shown that canine BMSCs can form neurosphere-like clumps and differentiate into neuron-like cells expressing neuronal markers [6, 8, 12, 13, 20, 24]. However, to the best of our knowledge, the differentiation of canine BMSCs into functional neurons has not yet been proven.…”

Isolation, characterization and immunomodulatory-associated gene transcription of Wharton’s jelly-derived multipotent mesenchymal stromal cells at different trimesters of cow pregnancy

“…For all procedures 2 × 10 5 cells/ml were submitted to osteogenic, chondrogenic, adipogenic and neurogenic differentiation according to the manufacturer's instructions (STEMPRO ® differentiation medium; Invitrogen). The neurogenic differentiation was adapted from previous studies (Oda et al 2013). For osteogenic/chondrogenic differentiation, 2 ml of STEMPRO ® osteogenic/chondrogenic differentiation medium was added to undifferentiated cultures comprising osteogenic and chondrogenic commercial inducers (STEMPRO ® ).…”

“…In order to verify adipocytes, Oil Red staining was performed (Cardoso et al 2012). The differentiation of bovine-derived WJMSC cells into neural-like cells followed the procedure described previously, with some modifications (Cardoso et al 2012;Oda et al 2013;Silva et al 2016). The neuronal differentiation was confirmed by immunofluorescence for GFAP and nestin cell markers, as described previously (Cardoso et al 2012).…”