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Isolation and culture method of amniotic-fluid-derived neural stem cells

A technique for the isolation and cultivation of neural stem cells, applied to nervous system cells, animal cells, vertebrate cells, etc., can solve problems such as unclear biological characteristics, and achieve the effect of strong three-dimensional sense, high success rate, and clear boundaries

Inactive Publication Date: 2011-10-05
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, research on amniotic fluid-derived stem cells is still in its infancy, and many of its biological properties are still unclear
So far, there is no report on the isolation and culture of amniotic fluid-derived neural stem cells (Amniotic Fluid-derived Neural Stem Cells, AFNS cells)

Method used

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  • Isolation and culture method of amniotic-fluid-derived neural stem cells
  • Isolation and culture method of amniotic-fluid-derived neural stem cells
  • Isolation and culture method of amniotic-fluid-derived neural stem cells

Examples

Experimental program
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Embodiment 1

[0050] Example 1 Separation, Culture and Induction of Differentiation of Porcine Amniotic Fluid-derived Neural Stem Cells

[0051] 1.1 Isolation and culture of porcine amniotic fluid-derived neural stem cells

[0052] 1) Isolation and culture of amniocytes

[0053] Fetal amniotic fluid was aseptically collected from pigs of 30 days of pregnancy, and the collected fetal amniotic fluid was successively filtered through 200 mesh and 400 mesh cell sieves, the filtrate was collected, and the amniotic fluid supernatant was collected after centrifugation at 1000r / min for 8min; then mixed culture medium ( Amniotic fluid supernatant and amniotic fluid cell culture medium at a volume ratio of 1:1) to resuspend the sediment (cell pellet) to prepare a cell suspension.

[0054] Inoculate 2mL of cell suspension into a 25cm2 adherent cell culture flask (or inoculate 6mL of cell suspension into a 75cm2 adherent cell culture flask), add 15mL (or 50mL) of mixed culture solution, and inoculate ...

Embodiment 2

[0086] Example 2 Isolation, culture and differentiation induction of goat water-derived neural stem cells

[0087] 2.1 Isolation and culture of goat water-derived neural stem cells

[0088] 1) Isolation and culture of amniocytes

[0089] Aseptically collect fetal amniotic fluid from Guanzhong dairy goats at the end of pregnancy, filter the collected fetal amniotic fluid through 200-mesh and 400-mesh cell sieves in turn, collect the filtrate, and collect the amniotic fluid supernatant after centrifugation at 1000r / min for 8 minutes; then use the mixed culture medium (amniotic fluid supernatant and amniotic fluid cell culture medium at a volume ratio of 1:2) to resuspend the precipitate (cell pellet) to prepare a cell suspension.

[0090] Inoculate 2mL of cell suspension into a 25cm2 adherent cell culture flask (or inoculate 6mL of cell suspension into a 75cm2 adherent cell culture flask), add 15mL (or 50mL) of mixed culture solution, and inoculate at 37°C, 5% CO 2Cultured in...

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Abstract

The present invention discloses an isolation and culture method of amniotic-fluid-derived neural stem cells. The method comprises the following steps of: firstly isolating to obtain amniotic fluid cells, then adding an isolation culture medium to the amniotic fluid cells, isolating obtained neural stem cells to grow and form neurospheres, digesting and isolating the neurospheres into single cells and carrying out subculture. The isolation and culture method of the amniotic-fluid-derived neural stem cells provided by the invention opens up a novel way for the isolation and culture of the neural stem cells, and has a high success rate in the isolation and culture the neural stem cells from the amniotic fluid. The isolated amniotic-fluid-derived neural stem cells can differentiate into astrocytes, neuronal cells and other functional cells after in-vitro induction, and have feasibility, effectiveness and the potential of self-renewal, proliferation, and differentiation.

Description

technical field [0001] The invention belongs to the technical field of neural stem cells, and relates to a method for separating and culturing amniotic fluid-derived neural stem cells. Background technique [0002] Neural stem cells (Neural Stem Cells, NSCs) refer to a class of cells with self-replication and renewal capabilities, high proliferation and multiple differentiation potentials, because they play an important role in the study of nervous system development, differentiation and treatment of nervous system diseases, so much attention. Anderson D J first proposed the concept of neural stem cells, and first confirmed the existence of neural stem cells through experiments, and then Cattaneo, Reynolds, etc. published papers on the discovery of neural stem cells and their related characteristics in Science and Nature respectively. The discovery of neural stem cells is a major development in neuroscience, marking the end of the theory that "central nerve cells cannot reg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797
Inventor 郑月茂贺小英刘军郑艳玲赵晓娥张涌
Owner NORTHWEST A & F UNIV
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