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Method for preparing sub totipotential stem cell by utilizing placenta lobular tissue

A technology of subtotipotent stem cells and placental leaflets, which is applied in the field of stem cell preparation, can solve problems such as slow proliferation speed and weak proliferation ability, and achieve the effects of short growth cycle, improved uniformity, and cost-saving preparation

Active Publication Date: 2013-09-04
台州恩源生物科技有限公司
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Problems solved by technology

The Chinese invention patent name is sub-totipotent stem cells, its preparation method and its use (application number CN200810240040.8) discloses a sub-totipotent stem cell, which is derived from a cut human umbilical cord or placenta, and the cell marker is CD151+OCT4+CD184-, It grows adherently in a culture vessel and can differentiate into human body, mesoderm and ectoderm tissues. Its preparation process needs to be expanded to more than four generations to obtain enough cells for the establishment of a sub-totipotent stem cell seed bank, and its proliferation ability is weak , a slower growth rate

Method used

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specific Embodiment

[0028] A method for preparing sub-pluripotent stem cells using placental lobule tissues specifically includes the following steps:

[0029] 1. Isolation of subtotipotent stem cells

[0030] 1.1 Placental pretreatment

[0031] After obtaining the consent of the parturients with normal prenatal examinations (no virus infection, no abnormal conditions during pregnancy) and signing the "Informed Consent", the placenta within 24 hours of being separated from the mother will be retrieved from the obstetric department of the hospital and enter the laboratory for backup;

[0032] Place the placenta in a sterile tempered dish in a hundred-level laminar flow room, add physiological saline to fully soak, remove the connective tissue at the junction of the placenta surface and the umbilical cord, cut the placental lobules and put them in a petri dish. 3 to 5 times of brine cleaning;

[0033] 1.2 Preparation of cell suspension

[0034] Cut the placental leaflet tissue into 0.5~1.5mm 3 Small pi...

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Abstract

The invention discloses a method for preparing a sub totipotential stem cell by a utilizing placenta lobular tissue. The method is characterized by comprising the following steps of: preparing cell suspension by utilizing the placenta lobular tissue after pretreating placenta, and preparing sub totipotential stem cell separating medium by utilizing the cell suspension; and carrying out primary culture and a second-generation culture on the sub totipotential stem cell separating medium, and collecting fully cast-off cells in the second-generation culture process to obtain the sub totipotential stem cell; and cooling and freezing in a programmed manner, taking out the frozen sample after cooling the temperature to -80 DEG C to -90.0 DEG C, and placing the frozen sample in a liquid-nitrogen storage tank for storing for a long time. The method for preparing the sub totipotential stem cell by utilizing the placenta lobular tissue has the advantages that the passage capacity can reach 20 generations, so that not only the capacity of differentiating towards bones, cartilages, fat and nerve cells can be achieved, but also the capacity of differentiating towards islet cells and epidermis cells be achieved; and the sub totipotential stem cell is uniform, stable, short in production period, simple to operate and low in cost.

Description

Technical field [0001] The present invention relates to the field of stem cell preparation, in particular to a method for preparing sub-pluripotent stem cells from placental tissue. Background technique [0002] Stem cell technology is one of the most popular technologies in life sciences today. Its research content covers almost all biomedical fields of life sciences. In addition to playing an important role in cell therapy, tissue / organ transplantation and gene therapy, it is also discovering new genes , Gene function analysis, developmental biology models and new drug development. Sub-totipotent stem cells are stem cells derived from mature placental tissues. They have high self-renewal ability and multi-directional differentiation potential. They are new seed cells for cell transplantation and tissue engineering and have broad application prospects. [0003] As an emerging and more promising project in the stem cell industry, subtotipotent stem cells have bright prospects, but...

Claims

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Application Information

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IPC IPC(8): C12N5/073
Inventor 陈正琳陈平李杰夏佳音
Owner 台州恩源生物科技有限公司
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