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Polysaccharide composition for promoting proliferation of human umbilical cord mesenchymal stem cells and application of polysaccharide composition

A quality stem cell and composition technology, applied in the field of biomedicine, can solve the problems of slow growth rate of stem cells, unstable culture effect, and reduced cell viability, and achieve the effect of remarkable effect, prevention of differentiation and promotion of proliferation ability.

Inactive Publication Date: 2018-07-06
南华生物医药股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, human umbilical cord mesenchymal stem cells have been widely used in various basic researches and clinical trials of disease treatment, but the current culture schemes used are different, and the culture effect is not stable. Stem cells often have slow growth and morphological changes leading to trait changes , decreased cell viability, etc.

Method used

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  • Polysaccharide composition for promoting proliferation of human umbilical cord mesenchymal stem cells and application of polysaccharide composition
  • Polysaccharide composition for promoting proliferation of human umbilical cord mesenchymal stem cells and application of polysaccharide composition
  • Polysaccharide composition for promoting proliferation of human umbilical cord mesenchymal stem cells and application of polysaccharide composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Isolation and purification of human umbilical cord mesenchymal stem cells: Wash the human umbilical cord three times with PBS buffer (containing penicillin and streptomycin) under sterile conditions, remove the adventitia and blood vessels of the umbilical cord, cut it into pieces and press the volume ratio of 1:3 Mix with low-sugar DMEM (Gibco) mixture containing trypsin (w / v 0.05%) and type IV collagenase (1.5 mg / mL), digest at 37°C for 1 h, and filter undigested tissue pieces through a 200-mesh filter. Then add hyaluronidase with a final concentration of 25ug / mL to digest for 10min, centrifuge at 100-200r / min for 10min, discard the supernatant, wash the precipitate once with low-sugar DMEM medium containing 10% FBS, and centrifuge at 100-200r / min Centrifuge for 10 minutes, remove the supernatant to obtain the precipitate, which is the primary umbilical cord mesenchymal stem cells; inoculate the obtained primary umbilical cord mesenchymal stem cells in the culture medi...

Embodiment 2

[0025] Ophiopogon japonicus polysaccharide and Atractylodes macrocephala polysaccharide composition and its preparation method: take 300 g of dried Ophiopogon japonicus and 100 g of Atractylodes macrocephala, pulverize and mix them evenly, add 4000 ml of pure water, boil for 2 hours; add 2000 ml of pure water again, and boil for 1 hour; filter to remove impurities, and the filtrate Concentrate in vacuo, the concentrated solution is alcohol-precipitated with 70% ethanol aqueous solution by volume, and the precipitate obtained by centrifugation is the composition of Ophiopogon japonicus polysaccharide and Atractylodes macrocephala polysaccharide.

Embodiment 3

[0027] Promoting effect of Ophiopogon japonicus polysaccharide and Atractylodes macrocephala polysaccharide composition on the proliferation of umbilical cord mesenchymal stem cells: Mix Ophiopogon japonicus polysaccharide and Atractylodes macrocephala polysaccharide composition (obtained in Example 2 or purchased Ophiopogon japonicus polysaccharide and Atractylodes macrocephala polysaccharide monomer in a ratio of 3:1 After filter sterilization), it was dissolved in DMEM / F12 medium at a ratio of 1%. The medium was used to culture umbilical cord mesenchymal stem cells (the mesenchymal stem cells were obtained from the umbilical cord according to the method in Example 1). The experimental groups were: medium control group (no polysaccharide composition added); medium + polysaccharide composition. After four days of culture, add serum-free DMEM / F12 medium to wash twice, then treat with MTT at a final concentration of 5 mg / mL, incubate at 37°C for about 4 hours, then carefully ab...

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Abstract

The invention discloses an ophiopogon polysaccharide and white atractylodes rhizome polysaccharide composition and preparation method thereof, and discloses application of the ophiopogon polysaccharide and white atractylodes rhizome polysaccharide composition in promotion of proliferation of human umbilical cord mesenchymal stem cells. The drug composition disclosed by the invention can enable mesenchymal stem cells to keep relatively high activity, substantially increase the proliferation speed and prevent differentiation of the cells. Application of the umbilical cord mesenchymal stem cellsprepared through the invention has important significance for a scientific research and clinical application.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to the preparation and application of a polysaccharide composition, in particular to the application of the Ophiopogon japonicus polysaccharide and Atractylodes macrocephala polysaccharide composition in promoting the proliferation of human umbilical cord mesenchymal stem cells. Background technique [0002] Human umbilical cord mesenchymal stem cells (Mesenchymal Stem Cells, MSCs) refer to a kind of cells with the characteristics of totipotent stem cells existing in the umbilical cord tissue of newborns. Differentiation, with multi-lineage differentiation potential, its morphology and biological characteristics are similar to those of bone marrow-derived MSCs (Br J Haematology, 2000, 109:235-242). Umbilical cord mesenchymal stem cells (MSCs) have high differentiation potential and can differentiate into osteoblasts, chondrocytes, adipocytes, nerve cells, liver cells, skeletal musc...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28
CPCC12N5/0665A61K35/28C12N2501/90
Inventor 向双林黄招琴彭海宁杨满君颜峰胡翔
Owner 南华生物医药股份有限公司
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