Aedating v4 1
Specific neuroregulatory molecules such as (D) cystatin C (Cys C), (E) glia-derived nexin (GDN), (F) galectin-1 (Gal-1) and (G) pigment epithelium-derived factor (PEDF) were found to be upregulated in the dynamic h BM-MSCs secretome (data are expressed as mean ± SEM, n=3). Trophic factor values were normalized to cell density (i.e. In the present work, we have expanded h BM-MSCs on microcarriers in computer-controlled stirred suspension bioreactor (Fig. This technology allows us to: (1) expand a large number of cells in one vessel, (2) monitor and ensure tight control of process variables such as p H, temperature and dissolved oxygen concentration (Fig.
pg (of each factor)) per 10,000 cells in each condition, respectively. 1C), and (3) develop a process, which allows for scale-up to larger bioreactor systems.
The implementation of computer-controlled suspension bioreactors has shown to be a viable route for the expansion of these cells to large numbers.
Numerous studies have evaluated the use of microcarriers and suspension bioreactors for the expansion of h MSCs.
Herein, we have shown that by expanding the h BM-MSCs in a computer-controlled suspension bioreactor system, similar cell doubling times and expression of h MSC surface antigens (Fig.
Indeed, we observed (on h NPCs-differentiated cultures with dynamic CM) a significant increase of NOTCH1 expression (F cells, which evidences their progenitor profile.
h BM-MSCs CM collected from static and dynamic culture conditions was able to significantly increase the survival and differentiation of h NPCs into (E, F) immature (DCX (p = 0.077) cells compared to the (E, I, M) static h BM-MSCs CM in the induction h NPC differentiation ((G, K, O); mean ± SEM., n = 3). 3F) resulted in a significantly higher number of DCX-expressing cells (p = 0.016) when compared to the static-CM group (Fig. After analyzing the co-expression of Ki-67 = 0.678) the number of differentiating neurons when compared to the Sham group (Fig. At the same time, although without statistical differences, the dynamic CM presented a positive trend when compared to the static CM (Fig. Similar effects were also obtained for astrocytic cells (Fig.
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Moreover, it has been demonstrated that h MSCs are able to secrete important neuroregulatory molecules such as brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), insulin growth factor 1 (IGF-1), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), transforming growth factor beta (TGF-β), glial-derived neurotrophic factor (GDNF), fibroblast growth factor 2 (FGF-2), stem cell factor (SCF), granulocyte colony-stimulating factor (G-CSF) and stromal cell-derived factor (SDF-1) both in vitro and in vivo.