Method for establishing human amniotic fluid stem cell line

A stem cell line and establishment method technology, applied in the field of cell line establishment, can solve the problems of reduced number, limited cell proliferation ability, low purity, etc., and achieve the effect of stable performance and good genetic traits

Inactive Publication Date: 2010-12-29
NORTHWEST A & F UNIV
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Problems solved by technology

However, the sorted cell population contains amniotic fluid-specific cells, epithelioid cells and other types of cells, and its purity is very low, and the proliferation ability of the cells is limited, and the number decreases after multiple passages, and only 10 passages can be expanded in vitro about
At present, no one has established a stable amniotic fluid stem cell line in China

Method used

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Embodiment Construction

[0037] The invention provides a method for establishing a human amniotic fluid stem cell line, the method comprising the following steps:

[0038]1. Amniotic fluid stem cell separation steps: collect amniotic fluid in the second and third trimester from the obstetrics and gynecology department of the hospital, record in detail the pregnancy time, amniotic fluid appearance, amniotic fluid volume, and the size of cell clumps after centrifugation, and store them at 4°C and bring them back to the laboratory. Filter with a 200-mesh filter to remove larger tissue fragments, then centrifuge at 1000r / min for 6min, discard the supernatant, and collect the cells. Then add PBS(-) solution containing 1% double antibody, mix well, treat at 37°C for 10min, then centrifuge, collect cells, repeat treatment with 1% double antibody PBS(-) solution 3 times.

[0039] 2. Amniotic fluid stem cell culture steps: 1) Add the amniotic fluid cells isolated in the previous step to the primary amniotic fl...

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Abstract

The invention relates to a method for establishing a human amniotic fluid stem cell line. The method comprises the following steps of: 1) performing isolated culture on primary amniotic fluid stem cells; 2) separating and purifying the primary amniotic fluid stem cells subjected to isolated culture; and 3) performing monoclonal screening and amplification culture on the separated amniotic fluid stem cells so as to establish the amniotic fluid stem cell line. The invention provides a rapid, effective and safe method for establishing the human amniotic fluid stem cell line, can realize long-term amplification of the human amniotic fluid stem cells in vitro, can provide a novel seed cell source for tissue engineering research, and makes the human amniotic fluid stem cells serve as seed cells for clinical application.

Description

technical field [0001] The invention relates to a method for establishing a cell line, in particular to a method for establishing a human amniotic fluid stem cell line. Background technique [0002] Stem cells have the ability of self-renewal, and under certain induction conditions, they can differentiate into cells with special functions. In the past ten years, the success of isolating and culturing embryonic stem cells (ES cells) has provided new opportunities for basic research and cell transplantation therapy. [0003] However, social and ethical issues restrict the research and application of ES cells. In addition, culturing ES cells in vitro faces great technical challenges. The culture of ES cells requires feeder layers and expensive cytokines, and genome mutations often occur during in vitro culture. More importantly, ES cells may form tumors after transplantation in vitro, and there is no solution yet. However, the number of adult stem cells such as bone marrow m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12R1/91
Inventor 王华岩陈帅王晗窦忠英
Owner NORTHWEST A & F UNIV
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