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Method for extracting and purifying multi-source stem cells from placenta and umbilical cord

A technology of umbilical cord blood stem cells and stem cells, which is applied in the field of simultaneous extraction and purification of stem cells from multiple sources, can solve problems such as transplant failure, endangering the life of patients, and excessive rejection, and achieve non-tumorogenicity and tumor-promoting, collection Easy to use, safe and reliable to use

Inactive Publication Date: 2013-09-25
周胜利
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  • Application Information

AI Technical Summary

Problems solved by technology

But chemotherapy does not distinguish between friend and foe. While killing cancer cells, it also kills normal hematopoietic stem cells, resulting in a lack of blood cells in the human body and endangering the patient's life.
In order to restore the hematopoietic function of the patient as soon as possible and save the patient's life, hematopoietic stem cell transfusion is required. However, if the immune markers of two people are too different, it will cause excessive rejection, making the transplantation fail and the patient dies.

Method used

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  • Method for extracting and purifying multi-source stem cells from placenta and umbilical cord
  • Method for extracting and purifying multi-source stem cells from placenta and umbilical cord
  • Method for extracting and purifying multi-source stem cells from placenta and umbilical cord

Examples

Experimental program
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Embodiment 1

[0070] Example 1: Collection of umbilical cord blood

[0071] see figure 1 , shows a culture of stem cell colonies derived from umbilical cord blood, each colony representing a hematopoietic stem cell.

[0072] Umbilical cord blood was collected by mechanical venipuncture, and the collected placenta came from local maternity hospitals or departments. First, after obtaining the consent of the pregnant woman or her immediate relatives to donate placental tissue, check all the test reports of the pregnant woman to confirm that there is no virus, syphilis and other blood-related viral infections, and then ask the pregnant woman in detail about her fertility, disease, heredity, infection, etc. Medical history, after everything was normal, after the pregnant woman gave birth, the placenta and cord blood were collected aseptically. After collection, the cord blood and placental tissue were attached with barcode labels and stored in a 4°C refrigerator. The storage time is generally...

Embodiment 2

[0073] Embodiment two: collect placental blood

[0074] see figure 2 , shows a colony culture of hematopoietic stem cells derived from placental blood. The figure shows the culture of stem cell colonies from placental blood. For the comparison of the same initial concentration of cells, the number of stem cell colonies from placental blood is far greater than that from umbilical cord blood at the same time.

[0075] Put the placenta after the umbilical cord blood has been taken in a high-pressure sterilized stainless steel container, insert a rubber tube connected with a syringe, infusion set or infusion pump into the umbilical cord artery, and enter the placenta through the vein, so that the residual blood in the placenta can flow out of the placenta Squeeze out and wash with a physiological buffer at the same time. The composition of the physiological buffer can be physiological saline, low molecular weight dextrose, phosphate-buffered saline or cell culture fluid, like 16...

Embodiment 3

[0076] Example 3: Collection of amniotic fluid and placental mesenchymal stem cells

[0077] see image 3 , the diagram shows the in vitro cell culture of placental mesenchymal stem cells after collection, concentration and purification, and the cells were attached after 24 hours of culture, showing fibrous cells.

[0078] There are two ways to remove the placenta tissue from healthy puerpera, one is for normal vaginal delivery and the other is for cesarean section. Free from contamination by bacteria and other pathogens.

[0079] In normal vaginal delivery, since the fetus and placenta must escape through the non-sterile vagina, which brings the possibility of microbial contamination, our experiments show that the possibility of microbial contamination of the extracted cells is greatly reduced.

[0080] The placenta is obtained by cesarean section under sterile environment and conditions or under aseptic conditions as far as possible.

[0081] 1) Collect the amniotic fluid...

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Abstract

The invention relates to a method for extracting and purifying multi-source stem cells from placenta and umbilical cord, the stem cells comprising cord blood hematopoietic stem cells, placenta blood hematopoietic stem cells, placenta amnion, amniotic fluid, chorion stem cells, and mesenchymal stem cells from placenta organization and umbilical cord organization. The method comprises: collecting amniotic fluid and cord blood by blood sampling device, then collecting placenta blood by a machinery step, and finally extracting, concentrating and purifying good-activity high-concentration large-number epidermal and mesenchymal stem cells from amniotic fluid, placenta amnion, chorion epidermal stem cell and placenta organization by using a machinery method, a combined enzyme digestion method and a monoclonal antibody-magnetic bead method. The stem cells are respectively carried out various detections, concentrated, purified and preserved, thereby a combined stem cell database is established, and the stem cells can be used for hematopoietic stem cell transplantation and cell therapy.

Description

technical field [0001] The invention relates to a method for extracting and purifying stem cells from placenta and umbilical cord, in particular to a method for simultaneously extracting and purifying stem cells from multiple sources from placenta and umbilical cord. Background technique [0002] Today in the 1990s, with the improvement of medical standards, many malignant infectious diseases and malnutrition diseases have gradually decreased or even disappeared. However, malignant tumors, genetic diseases, and immune system diseases are increasingly becoming the main diseases that involve a wide range of areas, cause serious harm, and are difficult to cure. For example, the annual incidence of malignant tumors in my country is 1.6 million, and there is a trend of increasing year by year; the main treatment methods are surgery, radiotherapy, and chemotherapy. Although the long-term survival rate of malignant tumors has increased with the improvement of these methods, most of...

Claims

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Application Information

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IPC IPC(8): C12N5/0735C12N5/0775C12N5/0789
Inventor 周胜利胡当玉周凝徐统乐胡晓辰
Owner 周胜利
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