Bone marrow mesenchymal stem cells (MSCs) have active abilities of self-replication and multidifferentiation. In recent years, a lot of studies have proved that MSCs can be induced and differentiated into nerve-like cells under certain conditions. Because of some advaced characteristics including sampling convenience, no immune rejection, high transfection rate and stable exogenous gene expression, MSCs will provide new way in treating disease of nervous system. In this article, the research progress of bone marrow mesenchymal stem cells differentiation into nerve-like cells induced by Traditional Chinese Medicine shall be discussed, and explore the research thinking guided by basis theory of TCM.
Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Differentiation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Neurons ; cytology ; drug effects

OBJECTIVETo set up a platform for phenotype-based primary screening of drug candidates promoting neuronal subtype differentiation in embryonic stem cells (ES) with light microscope.

METHODSHanging drop culture 4-/4+ method was employed to harvest the cells around embryoid body (EB) at differentiation endpoint. Morphological evaluation for neuron-like cells was performed with light microscope. Axons for more than three times of the length of the cell body were considered as neuron-like cells. The compound(s) that promote neuron-like cells was further evaluated. Icariin (ICA, 10(-6)mol/L) and Isobavachin (IBA, 10(-7)mol/L) were selected to screen the differentiation-promoting activity on ES cells. Immunofluorescence staining with specific antibodies (ChAT, GABA) was used to evaluate the neuron subtypes.

RESULTSThe cells treated with IBA showed neuron-like phenotype, but the cells treated with ICA did not exhibit the morphological changes. ES cells treated with IBA was further confirmed to be cholinergic and GABAergic neurons.

CONCLUSIONPhenotypic screening with light microscope for molecules promoting neuronal differentiation is an effective method with advantages of less labor and material consuming and time saving, and false-positive results derived from immunofluorescence can be avoided. The method confirms that IBA is able to facilitate ES cells differentiating into neuronal cells, including cholinergic neurons and GABAergic neurons.

Animals ; Cell Differentiation ; drug effects ; physiology ; Cell Line ; Drug Evaluation, Preclinical ; methods ; Embryoid Bodies ; cytology ; Embryonic Stem Cells ; cytology ; Mice ; Nerve Regeneration ; drug effects ; Neurons ; cytology ; Phenotype

Spinal cord injury is a serious disabling disease caused by a series of internal and external factors in the field of orthopaedics and neuroscience, which is a big problem for doctors all over the world. Lithium has been used to treat dipolar disorder for over 100 years. It has been reported that lithium is benefit for brain neuron. The treatment effect for spinal cord injury gets more and more attention. Researches indicate that lithium is benefit for spinal cord injury by protecting neuron,reducing after-injury inflammation increasing the produce and release of neurotrophins, stimulating neurogenesis, enhancing autophagy and inhibiting apoptosis. This article summaries advances in mechanism of treatment for spinal cord injury with lithium by reviewing and analyzing researches. Therapy combined with lithium has a good application prospect.
Animals ; Apoptosis ; drug effects ; Humans ; Lithium ; therapeutic use ; Neurons ; cytology ; drug effects ; Spinal Cord Injuries ; drug therapy ; physiopathology

OBJECTIVETo explore the neuroprotection and the impact on brain development of lithium, the effects of lithium salt on the growth and survival of primary cultured cerebrocortical neurons were studied.

METHODSThe technique of primary cultured cerebrocortical neurons of newborn rats with serum-free medium was established, and the growth and survival of neurons treated with different doses of lithium chloride (0.625, 1.250, 2.500, 5.000, 10.000 mmol/L) were observed. The length of neuronal synapse, cell viability by MTT reduction assay were also measured.

RESULTSThe neurons were brighter, germinated rapidly, the neuronal synapse lengthened markedly, and the neurons viability was also better after treated with lithium chloride. Among the five doses, 5.000 mmol/L had the best effect [(53.80 +/- 5.84) micro m, P < 0.01].

CONCLUSIONLithium chloride can promote the growth and survival of neurons.

Animals ; Animals, Newborn ; Cell Division ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Cerebral Cortex ; cytology ; Lithium ; pharmacology ; Neurons ; cytology ; drug effects ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Wistar

BACKGROUNDY-27632 is a specific inhibitor of Rho-associated coiled kinase (ROCK) and has been shown to promote the survival and induce the differentiation of a variety of cells types. However, the effects of Y-27632 on adult human adipose tissue-derived stem cells (ADSCs) are unclear. This study aimed to investigate the effects of Y-27632 on the neuronal-like differentiation of ADSCs.

METHODSADSCs were isolated from women undergoing plastic surgery and cultured. ADSCs were treated with different doses of Y-27632 and observed morphological changes under microscope. The expression of nestin, neuron specific enolase (NSE) and microtubule-associated protein-2 (MAP-2) in ADSCs treated with Y-27632 was detected by immunocytochemistry and Western blotting analysis.

RESULTSY-27632 had the potency to induce neuronal-like differentiation in ADSCs in a dose-dependent manner. Moreover, the differentiation induced by Y-27632 was recovered upon drug withdraw. ADSCs treated with Y-27632 expressed neuronal markers such as NSE, MAP-2 and nestin while untreated ADSCs did not express these markers.

CONCLUSIONSelective ROCK inhibitor Y-27632 could potentiate the neuronal-like differentiation of ADSCs, suggesting that Y-27632 could be utilized to induce the differentiation of ADSCs to neurons and facilitate the clinical application of ADSCs in tissue engineering.

Adipose Tissue ; cytology ; Adult ; Amides ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Female ; Humans ; Neurons ; cytology ; Pyridines ; pharmacology ; Stem Cells ; cytology ; drug effects

AIMTo establish model of differentiation of fetal liver stem cells induced by beta-ME + BHA into neural cells in vitro;

METHODSCD34+ cells from naturally aborted human fetal liver were isolated with MACS Kit, and cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS). After confluent more than 80%, the 5 passage cells were induced by 10(-3) mol/L beta-mercaptoethanol (beta-ME) and 2 x 10(-4) mol/L BHA for 24 hours, and washed with PBS, and then incubated in serum-free medium for 5 hours to 5 days. The characteristics of treated cells were assayed by immunocytochemistry staining analysis.

RESULTSCells treated by beta-ME+ BHA exhibited neuronal phenotype, and expressed neuronal specific proteins such as nestin, NeuN, TrnJ-1, and NF-M, which were not found in control cells. Statistic analysis showed that 81% cells were NeuN-positive, 75% cells TuJ-1-positive, 47% cells NF-M-positive, 90% cells NSE-positive.

CONCLUSIONbeta-ME + BHA can induce human fetal liver CD34+ cells to produce neuronal specific antigens and proteins in vitro and become neuronal cells. CD34+ cells from human fetal liver possess potentials of differentiation into neural cells.

Antigens, CD34 ; Butylated Hydroxyanisole ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Liver ; cytology ; embryology ; Mercaptoethanol ; pharmacology ; Neurons ; cytology

OBJECTIVETo investigate the effect of ginsenoside Rg1 (G-Rg1) on the morphology of the hippocampal neurons of rats with electrical hippocampal injuries and evaluate its protective effects on the learning and memory function.

METHODSForty female SD rats were randomly divided into G-Rg1 group, saline group, sham-operated group and G-Rg1+Sham operation group. Using the stereotactic apparatus, electrical hippocampal injury was induced, not in the two sham groups, by application of direct electrical current, followed by treatments with intragastric administration of G-Rg1 or saline for 14 consecutive days. The learning and memory function of the rats was assessed with Morris water maze test. The viability and arrangement of the hippocampal neurons and the number of Nissl bodies were observed after the treatments.

RESULTSTreatment with G-Rg1 significantly improved the learning and memory function of rats with electrical hippocampal injury. The viability of the hippocampal neurons showed no significant changes in the two sham-operated groups (P>0.05), and the number of Nissl bodies was much lower in saline group than in the other groups (P<0.05).

CONCLUSIONSG-Rg1 can improve the learning and memory function of rats with electrical hippocampal injury, the mechanism of which is probably associated with its protective effect on the hippocampal neurons against electrical injury.

Animals ; Female ; Ginsenosides ; pharmacology ; Hippocampus ; cytology ; drug effects ; pathology ; Maze Learning ; drug effects ; Memory ; drug effects ; Neurons ; drug effects ; Rats ; Rats, Sprague-Dawley

Effects of lysophosphatidic acid (LPA), an extracellular phospholipid signal, on the proliferation of rat embryonic neural stem cells (NSCs) and their differentiation into microtubule-associated protein 2 (MAP2)-positive and choline acetyltransferase (ChAT)-positive, i.e. cholinergic-committed neurons, were observed in vitro by [(3)H]-thymidine incorporation, immunocytochemistry, Western blot and other techniques. The results showed that: (1) Lower concentrations of LPA (0.01~1.0 mumol/L) dose-dependently enhanced the uptake of [(3)H]-thymidine by NSCs cultured in specific serum-free medium, indicating a significant promotive action of LPA on the proliferation of NSCs. (2) After fetal bovine serum which induces and commences the differentiation of NSCs, was used in the medium, the lower concentrations of LPA increased the percentages of both MAP2- and ChAT-immunoreactive neurons, with a peak at 0.1 mumol/L LPA in two cases. (3) The promotive effects of LPA on the differentiation of MAP2- and ChAT-positive neurons were also supported by the up-regulation of the expressions of both MAP2 and ChAT proteins detected by Western blot. (4) At the early phase of differentiation of NSCs, the cell migration and neurite extension were enhanced significantly by lower dosages of LPA under phase-contrast microscope. These results suggest that LPA within certain lower range of concentrations promotes the proliferation of NSCs and their differentiation into unspecific MAP2-positive and specific cholinergic-committed neurons, and also strengthens the migration and neurite extension of the newly-generated neuronal (and also glial as reported elsewhere) progenitors.
Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cholinergic Neurons ; cytology ; Embryonic Stem Cells ; drug effects ; Lysophospholipids ; pharmacology ; Neural Stem Cells ; drug effects ; Rats

Neuronal PC12 cells induced by nerve growth factor (NGF) have been considered to be postmitotic and lack the ability to divide. However, in this study, we not only detected DNA synthesis but also observed cell division in some morphologically differentiated neuronal PC12 cells bearing long neurites. More interestingly, in addition to the division of perikaryon, the neurites located on the division site of the cell membrane also divided into two parts and were allocated to the two daughter cells. These results demonstrate that the morphologically differentiated neuronal PC12 cells still retain the ability to divide. This is the first report that neuronal PC12 cells as well as their neurites can divide.
Animals ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; DNA Replication ; drug effects ; physiology ; Nerve Growth Factor ; pharmacology ; Neurites ; drug effects ; Neurons ; cytology ; PC12 Cells ; Rats

Animals ; Habenula ; cytology ; drug effects ; Male ; Microelectrodes ; Neurons ; drug effects ; physiology ; Pain ; Pregnanolone ; pharmacology ; Rats ; Rats, Wistar