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Preparation and application of human-stem-cell-secreted bioactive factor and lysis solution

A technology of biologically active factors and biologically active substances, applied in medical preparations containing active ingredients, embryonic cells, reproductive cells, etc. Low material purity and other problems, to achieve the effect of improving body immunity, strong proliferation and differentiation ability, and high yield

Inactive Publication Date: 2012-07-18
ANHUI HUIEN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the stem cell bioactive factors or substances used in practice have the following deficiencies: (1) direct extraction from tissues such as placenta, resulting in low purity of stem cell bioactive factors or stem cells; Stem cell culture expansion (for example, using fetal bovine serum) in culture medium, the extracted secretions inevitably contain animal-derived components and even specific viruses that exist in animals. If it is used in clinical or health care, there will be (3) Embryonic stem cells are not easy to control in the process of proliferation and differentiation, and are highly tumorigenic. The secreted substances extracted from such stem cells may contain factors that cause malignant transformation of normal stem cells, while the degree of differentiation of stem cells in bone marrow and other tissues Higher, began to express related human leukocyte antigens, direct application may produce immune rejection; (4) Stem cells contain rich biologically active substances, which play an important role in the proliferation and differentiation of stem cells, but in the process of proliferation and differentiation, these substances do not It is not secreted outside the cells, and there is no patented technology for extracting biologically active substances in mesenchymal stem cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1, preparation of umbilical cord tissue stem cells

[0015] Stem cells are extracted from the umbilical cord tissue of healthy people. The stem cells are mainly mesenchymal stem cells, and the obtained stem cells are cultured and expanded in vitro. After the primary cells were collected, they were subcultured at a ratio of 1:2. When the cells were about 80% confluent, they were digested and collected with trypsin, and then subcultured at the same ratio, up to the fifth generation, and collected within five generations ( Including the cells of the fifth generation), the stem cells collected at this time have stable biological properties and strong value-proliferating ability. At the same time, the waste culture medium replaced during the cultivation and expansion process is collected and stored in a -80°C refrigerator. The specific process is as follows:

[0016] ①Collect the umbilical cord of a healthy person;

[0017] ②Remove the inner blood vessels and ou...

Embodiment 2

[0025] Embodiment 2, placental tissue stem cell preparation

[0026] Stem cells are extracted from the placental tissue of healthy people. The stem cells are mainly mesenchymal stem cells, and the obtained stem cells are cultured and expanded in vitro. After the primary cells were collected, they were subcultured at a ratio of 1:2. When the cells were about 80% confluent, they were digested and collected with trypsin, and then subcultured at the same ratio, up to the fifth generation, and collected within five generations ( Including the cells of the fifth generation), the stem cells collected at this time have stable biological properties and strong value-proliferating ability. At the same time, the waste culture medium replaced during the cultivation and expansion process is collected and stored in a -80°C refrigerator. The specific process is as follows:

[0027]①Collection of placental subamniotic tissue or chorionic tissue from healthy people;

[0028] ②Repeatedly rinse...

Embodiment 3

[0036] Embodiment 3, preparation of bioactive factors secreted by umbilical cord or placental tissue stem cells

[0037] Melt the waste liquid collected during the stem cell culture and expansion process, mix, filter through a 0.25 micron filter, and then ultrafilter through a hollow fiber ultrafiltration column with a molecular weight cut-off of 50KD. Concentrate the ultrafiltered liquid to obtain rich stem cells. Composition of biologically active factors.

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Abstract

The invention discloses preparation and application of human-stem-cell-secreted bioactive factors and a lysis solution. The process comprises the following steps of: extracting stem cells of the umbilical cord or the placenta tissue of a health human person; performing in-vitro culture and amplification; and preparing high-purity and high-activity stem-cell-secreted bioactive factors and bioactive substances in the stem cell (namely, high-purity stem cell lysis solution) by the modern biological technologies of low-temperature separation, purification and the like. According to the invention, the bioactive factors or the substances do not contain any animal component, and can be developed into a product with a cell biological activity; after acting on the human body, the product can activate the stem cell of the organism and repair or replace damaged, lesion and old cells; and the product has the functions of regulating the cell metabolism function of the organism, enhancing the immunity of the organism, delaying the aging and the like. The human bioactive factors or the substances have great application prospect in the fields of medical science, healthcare and beauty.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to extracting stem cells from umbilical cord tissue or placental tissue of healthy people, culturing and amplifying them in vitro, and then extracting bioactive factors secreted during stem cell culture and amplifying and bioactive substances in stem cells and apply. Background technique [0002] Stem cells (stem cells) are a type of cell population with self-renewal, high proliferation and multi-directional differentiation potential, and are the "ancestor cells" and "universal cells" that form various tissues and organs of the human body. According to their developmental stage, they can be divided into embryonic stem cells and adult stem cells, and according to their differentiation potential, they can be further divided into totipotent stem cells, pluripotent stem cells and unipotent stem cells. [0003] Mesenchymal stem cells (MSCs) are a type of tissue stem cells with mu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/0735A61K35/44A61K35/50A61K8/98A61P37/04A61P39/06A61Q19/00A61Q19/08A61K35/51
Inventor 张正前
Owner ANHUI HUIEN BIOTECH
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