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A method for constructing human placental mesenchymal cells library which is suitable for clinical applicationc

A technology of human placenta and cells, applied in the medical field, can solve problems such as being unsuitable for clinical application, and the cell survival efficiency has yet to be proved.

Inactive Publication Date: 2012-08-08
北京雅联百得科贸有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has two disadvantages. One is that the cell population obtained by this method is all monocytes, containing a variety of cells including lymphocytes, macrophages and adipocytes. Such a population is not suitable for clinical application
The second is that the cells obtained by this method are directly used for cryopreservation without in vitro culture, and the cell survival efficiency has yet to be proved

Method used

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  • A method for constructing human placental mesenchymal cells library which is suitable for clinical applicationc
  • A method for constructing human placental mesenchymal cells library which is suitable for clinical applicationc
  • A method for constructing human placental mesenchymal cells library which is suitable for clinical applicationc

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0121] Example 1: Isolation of human placental amniotic stromal cells and human placental chorionic stromal cells

[0122] Under sterile conditions, approximately 20 g of fresh human placental tissue was dissected from the postpartum placenta. The tissue was stored in a 50 ml centrifuge tube containing 20 ml of DMEM (Invitrogen, Catalog No. 11885084) containing 1% human cord blood serum and 1% penicillin / streptomycin solution (Invitrogen, Catalog No. 15140122). ) culture medium. The placental tissue in the above protective solution was transferred to the cGMP laboratory within 30 minutes. Prior to processing, the tissue was washed three times in 1% penicillin / streptomycin in PBS (Invitrogen, Prod. No. 14040133).

[0123] The chorionic discs were dissected from the placental tissue using sterile surgical scissors, washed three times in PBS, and cut to approximately 1 mm 3 sized pieces were then digested at 37°C with a combination of 270 units per milliliter of collagenase ...

Embodiment 2

[0125] Example 2: Preparation of Autologous Umbilical Cord Blood Serum and Human Umbilical Cord Blood Serum for Placental Cell Growth

[0126] Cord blood from each placental cell donor was collected and processed separately in the following manner. The cord blood was collected after delivery using a 50 ml syringe with a 16G needle. Insert a needle into the umbilical vein of the placenta and draw cord blood from the umbilical vein into a syringe. The blood was then transferred to a 50 ml centrifuge tube without anticoagulant. Collect 30-40 mL of cord blood in each tube. The collected blood was transferred to the cGMP laboratory within 30 minutes.

[0127] 2.1 Preparation of autologous cord blood serum

[0128] Cord blood collected in a centrifuge tube without anticoagulant was coagulated for 45 minutes at 37°C for different donors, cooled in ice water for 30 minutes, and then centrifuged at 1000g at room temperature for 10 minutes. minute. Transfer the upper serum of ...

Embodiment 3

[0133] Example 3: In vitro culture of human placental stromal cells using human umbilical cord blood serum

[0134] The human placental amniotic membrane and placental chorionic stromal cells isolated according to the method of Example 1 were prepared at 1 × 10 per milliliter of culture medium. 6 The density of (one million) cells was dispersed in DMEM complete medium and the cells were transferred to 25 cm in a volume of 7.5 ml per flask 2 tissue culture flask. The components of the DMEM complete medium are: 89% DMEM, 10% human umbilical cord blood serum, and 1% penicillin / streptomycin solution. Place the flask containing this cell at 37°C with 5% CO 2 cultivated in the air. One week later, the medium was replaced with freshly prepared DMEM complete medium, and the culture was continued for another week. Cells were subcultured at the end of the second week and every 3-4 days thereafter, using freshly prepared DMEM complete medium for each passage. The method of each pa...

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Abstract

A method for treating human placenta cells samples and the human placental cells samples obtained by the method, a method for constructing human placental mesenchymal cells' library and the human placental mesenchymal cells' library obtained by the method, the uses of the obtained human placenta cells samples and the human placental mesenchymal cells' library in treating human dysfunction or diseases caused by cells injury or cells dysfunction, and the method for treating human dysfunction or diseases caused by cells injury or cells dysfunction are provided. A method for searching human placental cells in the library and a method for preparing human umbilical cord blood serum are also provided.

Description

technical field [0001] The present invention belongs to the field of medical technology. In particular, the present invention relates to a comprehensive method for the cultivation of human placental stromal cells suitable for clinical use and the establishment and management of a cell bank composed of said cells, including a method for protecting standard placental samples suitable for clinical use, Methods of culturing placental stromal cells and preparation of human autologous umbilical cord blood serum required for the implementation of these methods, and methods of digital registration management and retrieval of this cell bank. Background technique [0002] Human placental amniotic and chorionic stromal cells contain undifferentiated stem cells that can generate more of the same stem cells through cell proliferation in vitro, or can generate functional cells of multiple different cell lineages through cell differentiation in vitro. These two properties of placental ste...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N5/077A61K35/50A61P3/10A61P9/00A61P7/00A61P25/00A61P25/16A61P25/28
CPCA01N1/0221A01N1/0226A61K35/50A61P25/00A61P25/16A61P25/28A61P7/00A61P9/00A61P3/10
Inventor 杨银学魏军李玉奎王立斌刘婷马晓娜张广意
Owner 北京雅联百得科贸有限公司
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