| Mesenchymal stem cells (MSCs) and neural progenitor cells (NPCs) have been regarded for their clinical therapeutic potential for central nervous system (CNS) pathologies. Their potential utility is a result of their intrinsic ability to repair damaged tissues, deliver therapeutic proteins and migrate to sites of pathology within the brain. However, it remains unclear whether the CNS promotes any changes in these potential therapeutic cells, which would be critical to understand before clinical application. A major component of the CNS is cerebrospinal fluid (CSF). Therefore, the aim of this study was to evaluate the influence that human CSF has on the function of human adipose-derived MSCs (hAMSCs) and human fetal-derived NPCs (hfNPCs) in regards to cell proliferation, survival, and migration. This study may provide insight into improving the clinical efficacy of stem cells for the treatment of CNS pathologies. Human cerebrospinal fluid affects adipose-derived mesenchymal stem cells and fetal-derived neural progenitor cells on proliferation and survivalPurpose:To investigate the influence of human cerebrospinal fluid (H-CSF) on the proliferation and survival of human adipose-derived mesenchymal stem cells (hAMSCs) and human fetal-derived neural progenitor cells (hfNPCs).Method:The H-CSF samples were collected from non-cancer patients who underwent CSF diversion procedures for normal pressure hydrocephalus or congenital hydrocephalus. To determine the effects of human CSF on hAMSCs and hfNPCs, hAMSCs were cultured in100%MSC complete media (Ctrl),25%artificial CSF plus75%MSC complete media (A-CSF) or25%human CSF plus75%MSC complete media (H-CSF) respectively. Meanwhile, hfNPCs were cultured in100%NPC complete media (Ctrl),25%artificial CSF plus75%NPC complete media (A-CSF) or25%human CSF plus75%NPC complete media (H-CSF). The proliferation capacity of hAMSCs and hfNPCs were determined using MTT and Ki67immunofluorescence staining assays. Moreover, to determine the effect of human CSF on the viability of hAMSCs and hfNPCs, annexin V and propidium iodide (PI) were used to assess apoptosis.Results:MTT assays, Ki67staining and flow cytometry of annexin V and PI were performed to test these changes. hAMSCs cultured in H-CSF had a higher proliferation capacity and a larger number of Ki67positive cells when compared with hAMSCs cultured in the Ctrl and A-CSF (MTT assay, H-CSF vs. Ctrl:p<0.05; H-CSF vs. A-CSF:p<0.05; Ki67assay, H-CSF vs. Ctrl:p<0.05; H-CSF vs. A-CSF:p<0.05). Meanwhile, hAMSCs in the H-CSF group had a lower percentage of apoptosis when compared with the Ctrl and A-CSF group (Figure1D)(H-CSF vs. Ctrl:p<0.05; H-CSF vs. A-CSF:p<0.05). Moreover, the hfNPCs cultured in H-CSF exhibited a higher proliferation capacity and larger number of Ki67positive cells when compared with other groups (MTT assay, H-CSF vs. Ctrl:p<0.05; H-CSF vs. A-CSF:p<0.05; Ki67assay, H-CSF vs. Ctrl:p<0.05; H-CSF vs. A-CSF:p<0.05). The hfNPCs cultured in H-CSF also exhibited a higher survival rate compared with other groups (H-CSF vs. Ctrl:p=0.018; H-CSF vs. A-CSF:p=0.010). In addition, the hAMSCs cultured in50%human CSF had the highest proliferation capacity when compared with other groups (Ctrl,25%,50%,75%and100%human CSF). The hfNPCs cultured in25%human CSF exhibited the highest proliferation capacity compared with the other groups (Ctrl,25%,50%,75%and100%human CSF).Conclusions:These results suggest that human CSF can promote proliferation and decrease the rate of apoptosis for both hAMSCs and hfNPCs. Human CSF affects hAMSCs and hfNPCs on migration capacityPurpose:To investigate the influence of human CSF on the migration speed and distance of hAMSCs and hfNPCs.Method:In order to determine changes in the mobility of hAMSCs and hfNPCs in response to Ctrl, A-CSF and H-CSF, nano-ridges and grooves, constructed of transparent polyurethane acrylate, were used to evaluate cell migration speed and distance. Cell speed and distance were calculated with MATLAB. In order to analyze the influence of human CSF on migration, hAMSCs or hfNPCs were pre-cultured in the different culture media. Then, boyden transwell chambers were used to evaluate the migration capacity of hAMSCs and hfNPCs.Results:In order to test the mobility of hAMSCs and hfNPCs, cell migration was evaluated on nano-patterned surfaces and migration speed and travel distance was determined by MATLAB after culturing in Ctrl, A-CSF and H-CSF. The hAMSCs displayed an increased migration speed and distance migrated in H-CSF when compared to hAMSCs cultured in Ctrl and A-CSF (speed, H-CSF vs. Ctrl:p<0.05; H-CSF vs. A-CSF:p<0.05; distance, H-CSF vs. Ctrl: p<0.05; H-CSF vs. A-CSF:p<0.05). The hfNPCs in H-CSF also displayed significant differences in both speed and distance migrated when compared with other groups (speed, H-CSF vs. Ctrl:p<0.05; H-CSF vs. A-CSF:p<0.05; distance, H-CSF vs. Ctrl:p<0.05; H-CSF vs. A-CSF:p<0.05). Transwell migration assays were used to assess their migratory capacity. hAMSCs pre-cultured with H-CSF displayed a higher number of migrated cells compared with hAMSCs pre-cultured with A-CSF and Ctrl group (H-CSF vs. Ctrl:p<0.05; H-CSF vs. A-CSF:p<0.05). Meanwhile, the hfNPCs pre-cultured in H-CSF also displayed a higher number of migrated cells when compared to Ctrl and A-CSF group (H-CSF vs. Ctrl:p<0.05; H-CSF vs. A-CSF:p<0.05). Moreover, both hAMSCs and hfNPCs that were pre-treated in higher concentrations of H-CSF showed an enhanced migration capacity when compared to other groups.Conclusions:These results demonstrate that H-CSF can enhance the migratory capabilities of both hAMSCs and hfNPCs. Human CSF regulates characteristics of hAMSCs and hfNPCs through insulin-like growth factor1Purpose:To investigate the insulin-like growth factor1(IGF-1) in human CSF influence on the characteristics of hAMSCs and hfNPCs.Method:IGF-1was measured in human CSF samples. Then the IGF-1antibody was used as an inhibitor to test whether neutralization of the IGF-1in CSF could affect the functional characteristics of hAMSCs and hfNPCs. MTT assay, transwell assay, the flow cytometry of apoptosis and CXCR4expression were performed to quantify changes in viability, proliferation, and migration capacities of hAMSCs and hfNPCs.Results:An MTT assay was performed to analyze the proliferation of hAMSCs and hfNPCs. The hAMSCs cultured in H-CSF+IGF-1inhibitor exhibited a lower proliferation capacity when compared with the H-CSF group (H-CSF+IGF-1inhibitor vs. H-CSF:p<0.05). The IGF-1inhibitor also exhibited a similar effect on the hfNPCs when added to the H-CSF (H-CSF+IGF-1inhibitor vs. H-CSF: p<0.05). Flow cytometry was performed to quantify apoptosis of hAMSCs and hfNPCs. Both hAMSCs and hfNPCs in H-CSF+IGF-1inhibitor groups displayed an increase in the rate of apoptosis when compared to the groups cultured without the IGF-1inhibitor (hAMSCs, H-CSF+IGF-1inhibitor vs. H-CSF:p<0.05; hfNPCs, H-CSF+IGF-1inhibitor vs. H-CSF:p<0.05). Moreover, hAMSCs and hfNPCs cultured in H-CSF+IGF-1inhibitor decreased the migration capacity compared with the H-CSF group (hAMCSs, H-CSF+IGF-1inhibitor vs. H-CSF:p<0.05; hfNPCs, H-CSF+IGF-1inhibitor vs. H-CSF: p<0.05). In addition, When IGF-1inhibitor was added to the human CSF, the expression of CXCR4in hAMSCs and hfNPCs displayed a statistically significant decrease (hAMSCs, H-CSF+IGF-1inhibitor vs. H-CSF:p<0.05; hfNPCs, H-CSF+IGF-1inhibitorvs. H-CSF:p<0.05).Conclusions:These results suggest that IGF-1in human CSF plays an important role in enhancing the proliferation and maintaining the viability of hAMSCs and hfNPCs. Moreover, human CSF can promote the migration capacity of hAMSCs and hfNPCs; and is suggestive that this may be partly due to the presence of IGF-1. In addition, human CSF can stimulate the expression of CXCR4in both stem cell types. |
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