RETRACTED ARTICLE: Differentiation of Mesenchymal Stem Cells into Neural Stem Cells Using Cerebrospinal Fluid

Abstract

Optimization of a methodology for mesenchymal stem cells (MSCs) differentiation into neural stem cells (NSCs) using cerebrospinal fluid (CSF). MSCs were extracted from umbilical cord blood from healthy, full-term, newborn infants and from the bone marrow of patients. CSF was taken from healthy adult volunteers and patients. Four groups investigated were: A (n = 8) cord blood MSC induced with healthy volunteer CSF (control group); B (n = 7): patient MSCs induced with health volunteer CSF; Group C (n = 12): patient MSCs induced with their own CSF; group D (n = 6): cord blood MSCs induced with patient CSF. Following induction, cell differentiation state was examined using microscopy, flow cytometry, and immunohistochemistry. There were significantly more clinically applicable MSCs in Groups B and C than groups A and D (P < 0.05) and Group B had significantly more clinically applicable MSCs than group C (P < 0.05). The presence of NSCs was as with the MSCs. Group B had significantly more clinically applicable NSCs than all of the other groups. In addition, group B cells grew significantly faster than the other groups (P < 0.05). Upon CSF induction, MSCs differentiated into NSCs suitable for clinical treatment. The source of the MSCs and/or CSF influenced the number of NSCs produced and the NSC growth rate. Thus, the source of MSCs and CSF should be considered before initiating a stem cell clinical treatment.

In Vitro Differentiation of Bone Marrow Mesenchymal Stem Cells into Neuron-Like Cells by Cerebrospinal Fluid Improves Motor Function of Middle Cerebral Artery Occlusion Rats

“…Cytokines are commonly used as induction agents because they play a role in promoting stem cell proliferation and differentiation through their own receptors (20). We have demonstrated that hCSF-treated human BMSCs could successfully differentiate into neuron-like cells (10, 11). Three days of hCSF-induction prompts neurite growth, and the cultured cells gradually developed into a tapered, triangular, and irregular shape.…”

“…The soma on day 7 was similar to the dendrite and axon-like structures of astrocytes. Compared with cytokine-induced cells, those that were induced with hCSF were quicker to differentiate and which also occurred in higher proportions (10, 11). This has resolved numerous problems associated with the use of culture media and stimulating factors.…”

“…In brief, Bone marrow cells from rats weighting 100–150 g were harvested and introduced into T25 flask (BD Falcon™, Franklin lakes, NJ, USA) and cultured in a complete medium containing 10% fetal bovine serum (FBS, HyClone, Logan, UT, USA) and assorted by fluorescence-activated cell sorting (FACS), as described previously (10, 11). …”

“…At day 4 post-induction by hCSF, cells were digested with 0.25% trypsin and resuspended to a concentration of 1 × 10 7  cells/ml. Cell differentiation state was examined using microscopy, Western blot analysis, and fluorescent immunohistochemistry staining as described previously (10, 11). …”

“…Therefore, we theorized that BMSCs in vitro differentiation into more neuronal precursor cells than glia and fibroblasts before transplantation may represent a more effective therapeutic strategy. We previously successfully induced the differentiation of BMSCs into neuron-like cells using human cerebrospinal fluid (hCSF) (10, 11). In the present study, human CSF-treated rat BMSCs were injected stereotaxically into the brain of rats with focal cerebral ischemia caused by middle cerebral artery occlusion (MCAO).…”