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Insulin-like Growth Factor -1 Induces Differentiation of Human Umbilical Cord Blood-derived Mesenchymal Stem Cell to Oligodendrocytes-like Cells

Author: LinLing
Tutor: WangZuo
School: Fujian Medical
Course: Neurobiology
Keywords: Cord blood mesenchymal stem cells Oligodendrocytes Insulin-like growth factor 1
CLC: R329
Type: Master's thesis
Year: 2011
Downloads: 11
Quote: 0
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Abstract


【Objective】 cultured in vitro to obtain a sufficient number of umbilical cord blood-derived mesenchymal stem cells (mesenchymal stem cells, MSCs) and induced differentiation of insulin-like growth factor -1 (Insulin-like growth factor-1, IGF-1), neonatal rat oligodendrocytes? Ⅱ type star-shaped glial progenitor cells (oligodendrocyte? the type Ⅱ astrocyte, O2A progenitor cells, also known as oligodendrocyte progenitor cells) culture supernatant of co-culture methods to improve the efficiency of the induction of differentiation in more oligodendrocytes in vitro culture, and to provide an experimental basis for oligodendrocyte-related diseases. [Methods] 1. Collect the cord blood of newborns under sterile conditions, Ficon-HyPaque lymphocyte separation medium density gradient centrifugation, isolated mononuclear cells were cultured in vitro passage amplification, purification 3rd generation cord blood MSCs, surface antigen was measured by flow cytometry to Preadipocyte measured differentiation. Joint application to the differentiation of oligodendrocyte-like cells induced cord blood MSCs 2. BFGF, IGF-1, neonatal rat O2A progenitor cell culture supernatant by inverted fluorescence microscopy and Western blot analysis of cells successfully induced OL the quantitative detection of the positive cell count and Galc detecting neonatal rats O2A progenitor cell culture supernatant can improve the efficiency of IGF-1 induced. [Results] 1. Adherent cells available after the third passage uniform spindle cells, the growth of a certain direction, arranged in parallel or spiral-shaped growth, cell growth significantly faster, two weeks to up to 80% confluence, showed strong growth and passaging ability. Identification of flow cytometry not expressed or very weak expression of hematopoietic cell markers CD34, CD45, stably expressing mesenchymal cells associated surface antigen CD44 and CD105 in; Adipogenic agents may be induced to differentiate into adipocytes. 2. Induced approximately 10d, you can see the adherent cells showed different: visible undifferentiated cells with two symmetrical or three small protrusions oligodendrocyte precursor cells and round; approximately 3 weeks, visible cell projection increases, and can be connected to each other, showing oligodendrocyte morphology. Experimental group 3 (basal medium 10ng/mL bFGF 10ng/mL IGF-1) and negative control group (bFGF, IGF-1) the difference was statistically significant (P <0.05), compared to the rest of the group and negative control compared the difference was not statistically significant (P> 0.05). The experimental group A (10ng/mL IGF-1), the experimental group B (culture supernatant) Galc expression level of the difference was not statistically significant (P> 0.05); experimental group A, the experimental group C (culture supernatant 10ng / mL IGF-1) Galc express the amount of the difference was statistically significant (P <0.05); experimental group B, C Galc the expression of the difference was statistically significant (P <0.05). [Conclusion] 1 mononuclear cells obtained by density gradient centrifugation continuous subculture, the higher the purity of mesenchymal stem cells. 2 cord blood MSCs of bFGF, under the combined effects of IGF-1 to oligodendrocyte differentiation, but the efficiency of the induction of differentiation, and its physiological activity needs further testing. The efficiency of the impact of three different concentrations of inducing agent induced, select the appropriate inducing concentration is induced by the key to the success of this experiment show, 10ng/mL bFGF, 10ng/mL IGF-1 have good induction effect. Improve the efficiency of IGF-1 induced differentiation in the neonatal rat O2A progenitor cell culture supernatant.

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