Placenta stem cell bank construction method and placenta tissue resuscitation method

A technology of placental stem cells and placental tissue, which is applied in the field of construction of placental stem cell banks, can solve the problems that stem cells cannot be preserved, and achieve the effects of prolonging storage time, ensuring activity, and simple operation methods

Inactive Publication Date: 2015-04-01
天晴干细胞股份有限公司
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Problems solved by technology

This method only expands and cultures one kind of stem cells in t

Method used

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  • Placenta stem cell bank construction method and placenta tissue resuscitation method
  • Placenta stem cell bank construction method and placenta tissue resuscitation method
  • Placenta stem cell bank construction method and placenta tissue resuscitation method

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specific Embodiment approach 1

[0041] Specific embodiment one: a kind of placental stem cell bank construction method of the present embodiment, it is to carry out according to the following steps:

[0042] 1. Placenta collection: collect the complete placenta with 3-5cm umbilical cord left, and reserve it;

[0043] 2. Pretreatment of placental tissue: take at least 3 placental tissues from step 1 and use tissue scissors and forceps to carefully peel and collect them. The collected tissues are washed with normal saline for blood and cut to a size of 0.5-1mm. 3 Large and small, wash with normal saline for 2 to 3 times, wherein, after each washing, centrifuge at 1300rpm for 6 minutes to collect the precipitate and continue to the next wash; after washing, collect the precipitate for later use;

[0044] 3. Preparation of tissue block cryopreservation solution: 1mL of the cryopreservation solution is composed of 0.68-0.72mL DMEM, 0.07-0.12mL fetal bovine serum, 0.01-0.02mL L-glutamine and 0.2mL dimethyl sulfoxi...

specific Embodiment approach 2

[0048] Embodiment 2: This embodiment differs from Embodiment 1 in that the placental stem cell bank includes, but is not limited to, placental chorionic mesenchymal stem cells, placental amniotic mesenchymal stem cells, and placental decidua basal mesenchymal stem cells. and placental amniotic epithelial cells. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0049] Specific embodiment three: the difference between this embodiment and specific embodiment one is that the placenta collected in step one is washed with sterile physiological saline to wash the surface blood, then placed in a sterile biological sample sampling bag, and placed in a collection box at 4 to 25 ° C. Internal transportation, the collection box transportation process requires constant temperature control to prevent bumps, and the collected placenta must be processed within 72 hours. Others are the same as in the first embodiment.

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Abstract

The invention discloses a placenta stem cell bank construction method and a placenta tissue resuscitation method, and relates to placenta stem cell bank construction methods. The placenta stem cell bank construction method comprises the following steps: after non-bacterial cleaning treatment to placenta tissues of at least three parts, the steps of tissue peeling, further sterilizing, shearing to be thin, protecting using a refrigerant, programmed freezing, cryogenic temperature temporary storage, long term storage with nitrogen canisters, and the like are carried out, and the placenta tissue with biological activities can be preserved for a long term. The placenta tissue preserved through the method can be used for the construction of a stem cell bank, resuscitation, enzymic digestion, cultivation, expansion, quality inspection and the like are performed after selective tissue part resuscitation according to required cell types, so as to obtain placenta amniotic membrance epithelial cells, placenta amniotic membrance mesenchyme, placenta chorion mesenchymal stem cell, placenta decidua serotina mesenchymal stem cells and the like. The method provided by the invention has the advantages that operation steps are simplified, manual intervention errors are reduced, the efficiency is improved, more stem cell resources are preserved, and a basis is provided for further personalized stem cell customization.

Description

technical field [0001] The invention relates to a method for constructing a placental stem cell bank and a method for resuscitating placenta tissue. Background technique [0002] The placenta is the link for the delivery of various nutrients that the baby depends on for survival, and is composed of amniotic membrane, chorion and decidua basalis. The amnion and chorion are of fetal origin, while the decidua basalis is of maternal origin. In addition to playing an important role in fetal development, nutritional support and maintenance of tolerance, the placenta is also a body's stem cell reserve. [0003] There are many kinds of pluripotent stem cells in the placenta, and at least three types of placental stem cells are known. The first type of pluripotent stem cells in the placenta are mesenchymal stem cells (Mesenchymal stem cells, MSCs). Tissue cells such as liver cells. [0004] The characteristics of placental MSCs that can suppress immunity and promote hematopoietic...

Claims

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Application Information

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IPC IPC(8): C40B50/06C12N5/0735
Inventor 张怡王灵娟付寅生刘艳青
Owner 天晴干细胞股份有限公司
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