| The research and application of stem cell, a burgeoning subject, is one of the most attracting fields in the life sciences within the recent decades. There is a spreading application prospect of this new subject to cytotherapy, tissue and organ renovation, growth biology and pharmacy. It brings about new cut-in point to the research of Traditional Chinese Medicine as well as new challenge to the modernization of Traditional Chinese Medicine research.Mesenchymal stem cells (MSCs) are one of the focuses of research for the present. In addition to hematopoietic stem cell, there are undifferentiated mesenchymal stem cells in marrow, which come from mesoblast. It is traditionally acknowledged that the function of mesenchymal stem cells is to help construct-microenvironment for the subsistence and differentiation of hematopoietic stem cells. Recent research discovered that it has potential faculty of differentiation toward diversity of cell lines. It is prone to be obtained from the acceptor, and there is no immune response after replantation; It is prone to be isolated, cultured, and amplified in vitro, and is able to remain the undifferentiating state for a long time; It also possesses high in vitro gene transfer rate and stability of efficiently expression to extraneous gene. Hence, MSC is not only the wonderful seeded cells for tissue engineer, but also the ideal model for the research of proliferation and differentiation principles.On the basis of the fundamental theory of Traditional Chinese Medicine, Plastrum Testudinis is likely to control the proliferation and differentiationof MSC through the adjustment of Bone Morphogenetic Protein 4(BMP4) expression. It is acknowledged in the Traditional Chinese Medicine that kidney is in charge of growth, bone and marrow forming. The traits of Plastrum Testudinis are saltytaste, sweet flavour, and calm. It has the efficacy of strengthening tendons and bones as well as nourishing YIN(feminine) and kidney. Research discovered that Plastrum Testudinis cultured in vitro can promote the proliferation of MSC and guide MSC. to differentiate to be nerve cells and Osteogenesis. BMP4 is growth promoting, and an important bone-forming and nerve-growth inducement factor. Hence, the proliferation-promotion and differentiation-promotion of Plastrum Testudinis to MSC is likely related to BMP4 closely.From the perspectives of Morphology and molecular biology, etc, the research examined the influence of Plastrum Testudinis serum on BMP4mRNA and BMP4 in MSC qualitatively and quantitatively through transfer and interpretation with the adoption of experiment technic of immune cell chemics, in situ hybridization and real-time fluorescence PCR etc.The research has accomplished primary culture of MSC and established the cell system of MSC. With the combination of density gradient centrifugation and screening nonadherent cells, MSCs of high purity can be obtained. The purity of MSCs could be ascertained with the positive identification of double criteria of CD44, or CD54 and Brdu, and negative identification of CD34 and CD45 coloration.The research found that Seropharmacological Plastrum Testudinis could promote proliferation of MSCs. With the technology of MTT and Brdu marker and methods of PCNA immune cells chemistry applied to be the guideline of MSC evaluation, the experiment turned out that Seropharmacological Plastrum Testudinis can promote proliferation of MSCs and this function is dose-dependentThe outcome of the research indicated that Seropharmacological Plastrum Testudinis can induce MSC highly express BMP4mRNA and BMP4, and dosage groups of Seropharmacological Plastrum Testudinis are does-dependent. BMP4 protein and positive mRNA mainly lies in small MSC cells with great multiplication, which indicates that BMP4 is likely to functionin the early multiplication of cells.Under the instruction of "the kidney being in charge of development,... |
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