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People by regulating cytokine change the direction of the NSC differentiation

1992, Engel et al, first isolated from the adult rat striatum in vitro sustained proliferation of the cell population of the differentiation potential of the neurons and astrocytes, which indicates that the brains of adult mice having NSC characteristicscell populations. In 1998, Eriksson confirmed that the adult brain is also the NSC, to have a new understanding of mechanisms of nervous system regeneration and treatment of diseases of the nervous system. Generally think the NSC is mainly present in the mammalian brain, the hippocampus, striatum, cerebellar hemisphere and third ventricle wall as well as the lateral ventricle wall and other areas. In this study, these parts of the rat brain tissue isolation, cultured cells from the morphological characteristics of to cell chemical properties are in line with the characteristics of stem cells, which is consistent with the literature. The nestin protein specific markers are currently widely used for identification NSC its expression began neurulation, gradually disappear after the start of the neuronal migration and differentiation, and with the completion of the differentiation of nerve cells and stops the expression. Different cytokines play a continuous role in NSC proliferation, differentiation process, and ultimately the formation of the various types of cells of the nervous system (except microglia). People by regulating cytokine change the direction of the NSC differentiation, but also want can NSC induced to differentiate into a specific cell type.

 

Bone morphogenetic protein 4 on the stem cells of the rat brain choline induced differentiation effect

Observe the bone morphogenetic protein 4 (BMP4) isolated primary cultured rat brain stem cells (NSC) differentiation induced by cholinergic effect. From 2-month-old rat hippocampus, striatum region isolated cells were cultured in DMEM/F12 medium containing EGF and bFGF, to Nestin (of nestin in parallel morphological characteristics of the cells in the light microscope,) cytochemical staining. After 24h, a switch of BMP4-containing culture medium, the culture was continued for 7 or 8 days. Morphological changes of the cells in the light microscope, parallel choline acetyltransferase (choline acetyltransferase staining of ChAT) and nestin double-labeled immune cells. The results showed that, plus BMP4 cultured for 7 or 8 days, the light microscope to see 34% of the cultured cells with the morphological characteristics of neurons. The immunocytochemical staining visible ChAT-positive cells and nestin-positive cells coexist ChAT-positive cells accounted for 28% of nestin-positive cells accounted for 38%. In short, in the culture broth was added the BMP4 can induce NSC differentiation into cells having a cholinergic properties

    Experiments show that the natural differentiation of neural stem cells (neural stemcell, NSC) is not induced by neuron C-amino butyric acid (C-amino-butyricacid, GABA) to neurons vast majority of spontaneous differentiation of other phenotypic neurons are very rare. Cholinergic neurons is an important class of neurons in the nervous system involved in learning, memory and other important functions. 2000 Lgnacio and other reports, in cultured neuronal cells, bone morphogenetic occurrence protein 9 (bonemorphogeneticproteins-9, BMP9) directly induced encoding choline acetyl transfer enzymes and acetyl choline transporter capsule gene's expression and raised acetylcholine in the synthesis, to intraventricular injection the BMP9 increase acetylcholine levels in the living body, which shows BMP9 is cholinergic differentiation factor of the central nervous system. The BMP family BMP4 also has the same effect. However, with BMP4 the NSC culture cholinergic-induced differentiation has not been reported. To observe the culture medium used in this experiment will BMP4 NSC's cholinergic induced differentiation effects.

 

we fracture repair process a BMP4mRNA Expression and localization of detected using BMP4cDNA probe

In this experiment, we fracture repair process a BMP4mRNA Expression and localization of detected using BMP4cDNA probe, in order to investigate their role in fracture healing. It was found that: (1) not detected in normal bone tissue and soft tissue and BMP4mRNA expression; (2) early fracture (fracture after 12 to 72 hours) that a BMP4mRNA expression; (3) BMP4 gene mainly in early fracture hematoma emerging within the cell and fracture the surrounding soft tissue, expressed in mesenchymal cells. Consistent with the the Nakase other reported results. Bone marrow stromal cells may be derived from these hematoma, osteoblast potential of stem cells, i.e. directed into the bone precursor cells, the emerging positive cells in muscle tissue compared can induce osteogenic precursor cells, i.e. mesenchymal cells.

Trauma to activate the BMP4 gene's expression and showed some regional, that is limited to the fracture callus formation around the area of soft tissue. Topical application of exogenous recombinant expression product BMP4 able to induce new bone formation in normal soft tissue ectopic, so they think, local BMP4 gene expression of the fracture hematoma the osteogenic ability is the very important factor in the process of fracture healing callus formation also shows that the fracture hematoma and soft tissue has a very important position in the process of fracture healing. BMP4 gene expression is also reported in the literature in embryonic tissue, organs and embryonic bone, endochondral bone repair fractures embryonic osteogenesis reproduction, BMP4 only bone injury repair related, are more likely to tissues and organs and embryos related.

 

Histological changes in the process of fracture healing is a complex and involve a number of different cell activities

Histological changes in the process of fracture healing is a complex and involve a number of different cell activities. Fracture occurs, the presence of the different stages of osteoblast and osteoclast number and their synthetic cell interstitial far can not meet the demand of the fracture healing. Local cell regulation mechanisms (including precursor cells, mesenchymal cells, other synergy cells, capillaries, lymphatic, nerve growth factor autocrine and paracrine) to produce a sufficient number of osteoblasts and osteoclasts.

With the development of recombinant DNA technology, making it possible therapeutic areas. Such as growth factors and some other regulatory peptides for the regulation of cell metabolism, division and differentiation in the bone healing process. Part in the process of fracture healing growth factors and cytokines involved in the formation of bone cartilage has been reported in the literature.

 

Demonstration and Localization of Bone Morphogenetic Protein( BMP) in Fracture Healing

Post-traumatic bone tissue regeneration and repair is very complete, and its repair is often no scar residue. The reason why there is such a perfect bone tissue healing ability, because the memory of bone tissue factor in successful bone, bone growth factors. Bone morphogenetic proteins (bone morphogenetic pro-tein, BMP) because of its emphasis on direct induction of soft tissue into bone. Application BMP4cDNA probe to detect the fracture healing process foreign callus localization and distribution of BMP4 gene expression to explore the BMP4 gene expression outside in closed fracture healing callus formation. 64 healthy SD rats with closed tibial fractures animal models. 12 hours after fracture, 1,3,5,7,9,14 and 28 days after operation. Drawn underwent cryostat, the digoxigenin labeled BMP4cDNA probe situ hybridization. Rat fracture after 12 hours to three days, the detection of fractures around hematoma within cells and muscle emerging mesenchymal cells BMP4mRNA expression as a positive signal. Show that The trauma activates expression of BMP4mRNA, and was regional involved in the repair of fractures, the fracture hematoma and soft tissue has a very important role in the process of fracture healing.