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Method for establishing and detecting amniotic fluid stem cell line of pig

A technology of stem cell line and establishment method, which is applied in the field of establishment of cell lines to achieve the effects of stable performance, low tumorigenicity and good hereditary traits

Inactive Publication Date: 2013-07-31
NORTHWEST A & F UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Domestic research on amniotic fluid stem cells has just started, and there are still a lot of gaps. There are only sporadic reports on the isolation and culture of human amniotic fluid cells, research progress, etc.

Method used

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  • Method for establishing and detecting amniotic fluid stem cell line of pig
  • Method for establishing and detecting amniotic fluid stem cell line of pig
  • Method for establishing and detecting amniotic fluid stem cell line of pig

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Embodiment Construction

[0052] The invention provides a method for establishing a porcine amniotic fluid stem cell line, the method comprising the following steps:

[0053] 1. Primary isolation and culture of pig amniotic fluid stem cells, and analysis of which period amniotic fluid cells are the best: 1) Collect the amniotic fluid of pregnant sows from the slaughterhouse, and bring them back to the laboratory together with the uterus. The uterus is opened aseptically first, the amniotic fluid completely wrapped by the placenta is separated, the size of the fetus is measured and recorded in detail, and then the amniotic fluid is extracted with a syringe. 2) Filter with a 100-mesh filter to remove larger tissue fragments, then centrifuge at 2000r / min for 10min, discard the supernatant, and collect the cells. 3) Add the collected cells to a PBS solution containing 5% serum + 5% double antibody, mix well, and place at 38°C, 5% CO 2 Treat in the incubator for 10 minutes, then centrifuge to collect the c...

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Abstract

The invention provides a method for establishing an amniotic fluid stem cell line of a pig, which comprises the following steps: (1) carrying out primary separation culture on the amniotic fluid stem cell; (2) mechanically separating and purifying the primary amniotic fluid stem cell subjected to the separation culture; and (3) detecting and culturing the separated and purified amniotic fluid stem cell and establishing the amniotic fluid stem cell line of the pig. The invention provides a method of a quick, effective, safe and separated amniotic fluid stem cell, can provide a seed cell sourcefor researching tissue engineering and is expected to be used for research, production and application of the seed cell.

Description

technical field [0001] The invention relates to a method for establishing a cell line, in particular to a method for establishing a porcine amniotic fluid stem cell line. Background technique [0002] As research on embryonic stem cells is bound and limited by ethical and moral issues, many researchers are trying to find new sources of stem cells. In 2003, Prusa et al. found OCT4-positive cells in amniotic fluid, indicating that pluripotent stem cells may exist in amniotic fluid. In 2007, Anthony et al. reported in the journal Nature Biotechnology that they had found a new source of stem cells that was easy to obtain and had a strong ability to proliferate. By screening and isolating CD117-positive cells in amniotic fluid in the second trimester, the researchers obtained a small number of cells with embryonic stem cell characteristics. They named this type of cells amniotic fluid stem cells (Amniotic Fluid Stem Cells, AFS cells), including fetal skin, amniotic membrane of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12R1/91
Inventor 王华岩卢智娟成德张淑金高志敏
Owner NORTHWEST A & F UNIV
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