Canine Adipose Tissue-derived Stem Cells Research Articles

Comparison of the Efficacy of Surgical Decompression Alone and Combined With Canine Adipose Tissue-Derived Stem Cell Transplantation in Dogs With Acute Thoracolumbar Disk Disease and Spinal Cord Injury.

Paraparesis and paraplegia are common conditions in dogs, most often caused by a disc herniation in the thoracolumbar spinal segments (T3-L3), which is a neurological emergency. Surgical decompression should be performed as soon as possible when spinal compression is revealed by myelography, computed tomography, or magnetic resonance imaging. Mesenchymal stem-cell therapy is a promising adjunct treatment for spinal cord injury. This study sought to compare the effects of surgical decompression alone and combined with an allogeneic transplantation of canine adipose tissue-derived mesenchymal stem cells (cAd-MSCs) in the treatment of dogs with acute paraplegia. Twenty-two adult dogs of different breeds with acute paraplegia resulting from a Hansen type I disc herniation in the thoracolumbar region (T3-L3) were evaluated using computed tomography. All dogs had grade IV or V lesions and underwent surgery within 7 days after symptom onset. They were randomly assigned into two groups, 11 dogs in each. The dogs in Group I underwent hemilaminectomy, and those in Group II underwent hemilaminectomy and cAd-MSC epidural transplantation. In both groups, all dogs with grade IV lesions recovered locomotion. The median locomotion recovery period was 7 days for Group II and 21 days for Group I, and this difference was statistically significant (p < 0.05). Moreover, the median length of hospitalization after the surgery was statistically different between the two groups (Group I, 4 days; Group II, 3 days; p < 0.05). There were no statistically significant between-group differences regarding the number of animals with grade IV or V lesions that recovered locomotion and nociception. In conclusion, compared with surgical decompression alone, the use of epidural cAd-MSC transplantation with surgical decompression may contribute to faster locomotor recovery in dogs with acute paraplegia and reduce the length of post-surgery hospitalization.

Valproic Acid, a Histone Deacetylase Inhibitor, Decreases Proliferation of and Induces Specific Neurogenic Differentiation of Canine Adipose Tissue-Derived Stem Cells

ABSTRACTAdipose tissue-derived stem cells (ADSCs) isolated from adult tissue have pluripotent differentiation and self-renewal capability. The tissue source of ADSCs can be obtained in large quantities and with low risks, thus highlighting the advantages of ADSCs in clinical applications. Valproic acid (VPA) is a widely used antiepileptic drug, which has recently been reported to affect ADSC differentiation in mice and rats; however, few studies have been performed on dogs. We aimed to examine the in vitro effect of VPA on canine ADSCs. Three days of pretreatment with VPA decreased the proliferation of ADSCs in a dose-dependent manner; VPA concentrations of 4 mM and above inhibited the proliferation of ADSCs. In parallel, VPA increased p16 and p21 mRNA expression, suggesting that VPA attenuated the proliferative activity of ADSCs by activating p16 and p21. Furthermore, the effects of VPA on adipogenic, osteogenic or neurogenic differentiation were investigated morphologically. VPA pretreatment markedly promoted neurogenic differentiation, but suppressed the accumulation of lipid droplets and calcium depositions. These modifications of ADSCs by VPA were associated with a particular gene expression profile, viz., an increase in neuronal markers, that is, NSE, TUBB3 and MAP2, a decrease in the adipogenic marker, LPL, but no changes in osteogenic markers, as estimated by reverse transcription-PCR analysis. These results suggested that VPA is a specific inducer of neurogenic differentiation of canine ADSCs and is a useful tool for studying the interaction between chromatin structure and cell fate determination.

Adipose tissue-derived stem cells in oral mucosa tissue engineering: Enhanced migration and proliferation in co-culture with oral keratinocytes in vitro

Tissue-engineered oral mucosa holds a great prospect in urethroplasty and adipose tissue-derived stem cells (ADSCs) may play an important role in this field. In this research, canine oral keratinocytes (OKs) and ADSCs were harvested and cultured in vitro. The affinity between the two cell lines was evaluated by analyzing their migration and proliferation patterns in a co-culture environment. The results demonstrate that both canine ADSCs and OKs showed improved migration in the presence of the other cell line as a co-culture when compared to monoculture. Further, conditioned medium using the supernatant of one cell line accelerated the other cell line’s proliferation rate. Hence, it was concluded that the affinity between OKs and ADSCs was fitting; the presence of ADSCs accelerated the migration and proliferation of OKs in vitro. These results indicate that it is practical to use ADSCs and OKs to construct a tissue-engineered oral mucosa, since the presence of the former could activate the latter in vitro, maybe even in vivo. This may help to build tissue-engineered oral mucosa, which may be a new method for urethroplasty. Key words : Urethroplasty, adipose tissue-derived stem cells, oral keratinocytes, tissue engineering.

Role of laminin-111 in neurotrophin-3 production of canine adipose-derived stem cells: Involvement of Akt, mTOR, and p70S6K

Extracellular matrix (ECM) components play an important role in the regulation and maintenance of neural stem cells (NSCs). Laminin, an ECM component, is a key factor in promoting axonal regeneration and differentiation of NSCs. Since NSCs cannot be easily harvested with low morbidity, adipose-derived stem cells have been suggested for therapeutic applications of neural tissue damage. Therefore, the potential of laminin-111 to enhance the production of neurotrophin-3 (NT3) and its related signal pathways from canine adipose tissue-derived stem cells (cADSCs) was investigated. Laminin-111 enhanced NT3 production in neural induction medium (NIM). Treatment of NIM or laminin-111 on cADSCs distinctively changed integrin β1 mRNA and protein expression levels. In addition, laminin-111-induced Akt phosphorylation was inhibited by integrin β1 small interfering RNA (siRNA) and PI3K inhibitors (LY294002 and wortmannin). Furthermore, increased phosphorylations of mTOR and p70S6K by laminin-111 were blocked by inhibitors or specific siRNA, respectively. Moreover, laminin-111-induced NT3 production was blocked by these inhibitors. In experiments to induce the differentiation of cADSCs, laminin-111 increased the expression of neuronal markers β 3 tubulin, MAP2, and NeuN, and decreased the expression of the NSC markers nestin and vimentin. In conclusion, laminin-111 increases NT3 production through Akt, mTOR, and p70S6K pathways via integrin β1 in cADSCs cultured in NIM.