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Human-derived stem cell bioactive substance, its preparation and application

A technology of stem cell biology and active substances, applied in the field of preparation of biological active substances, can solve the problems of trauma, limited sources, high virus infection rate and other problems in collection, and achieve the effect of high yield and large comprehensive effect

Inactive Publication Date: 2012-05-23
张正前
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, MSCs are mainly derived from adult bone marrow, but the number and proliferation and differentiation potential of adult bone marrow MSCs decrease with the age of the donor, the virus infection rate is high, and the collection is traumatic, which limits its source

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1, preparation of umbilical cord tissue stem cells

[0014] Stem cells are extracted from the umbilical cord tissue of healthy people. The stem cells obtained from the umbilical cord tissue include but not limited to mesenchymal stem cells, and the obtained stem cells are cultured and expanded in vitro. After the primary cells are collected, they are subcultured at a ratio of 1:2. When the cells are about 80% confluent, they are digested and collected with trypsin, and subcultured at the same ratio. The maximum subculture is up to the fifth generation, and the cells within the fifth generation are collected. cell. The stem cells collected at this time have stable biological properties and strong value-added ability. The specific process is as follows:

[0015] ①Collect the umbilical cord of a healthy person;

[0016] ②Remove the inner blood vessels and outer membrane of the umbilical cord;

[0017] ③Cut the umbilical cord tissue into 1~2mm 3 Small pieces...

Embodiment 2

[0024] Embodiment 2, umbilical cord blood stem cell preparation

[0025] Nucleated cells are extracted from the umbilical cord blood of healthy people. The nucleated cells include but not limited to hematopoietic stem cells and mesenchymal stem cells. Using this kind of cells as the starting cells, they are inoculated on the trophoblast made of umbilical cord mesenchymal stem cells, and used Specific medium (this specific medium DMEM / F12: RPMI1640 mixed at 1:1, additionally containing 10% FBS and 10% umbilical cord blood plasma, pH = 7.0-7.4) for culture and expansion, and cultured for up to 10-11 days, Collect cells. Hematopoietic stem cells are the most abundant in the cells collected at this time. The cycle of single culture and expansion of hematopoietic stem cells is 10-11 days. The specific process is as follows:

[0026] ①Collection of umbilical cord blood from healthy people;

[0027] ② Add 5% hydroxyethyl starch (HES) at a volume ratio of 5:1 (cord blood: hydroxye...

Embodiment 3

[0036] Embodiment 3, preparation of umbilical cord and umbilical cord blood stem cell bioactive substance composition

[0037] Combine the umbilical cord blood-derived stem cells obtained in vitro with the umbilical cord-derived stem cells at a ratio of 1:5 to 1:10, and adjust the cell concentration to 1×10 7 ~1×10 8 Each / ml is sealed in a cryopreservation tube, and the ultra-low temperature liquid nitrogen is repeatedly frozen and thawed 4 to 5 times until the cells are broken, and then the high-purity stem cell bioactive substance group composition can be obtained through low-temperature separation, purification and other modern biotechnological means .

[0038] The bioactive substance group mainly comes from mesenchymal stem cells and hematopoietic stem cells, including but not limited to stem cell growth factor (SCF), nerve growth factor (NGF), stromal cell-derived growth factor (SDF), vascular endothelial cell growth factor (VEGF ), basic fibroblast growth factor (bFGF)...

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PUM

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Abstract

The invention discloses a human-derived stem cell bioactive substance, its preparation and application. The method comprises: first extracting healthy human-derived stem cells, which are then subjected to in vitro culture and amplification, collecting stem cells of specific phases, and utilizing modern ultralow temperature as well as other biological technologies to prepare a stem cell bioactive substance group composition. Mainly coming from mesenchymal stem cells and hematopoietic stem cells, the stem cell substance group can be developed into products with a cytobiological activity function. When applied on human bodies, the products can activate body stem cells, repair or substitute damaged, pathological and aging cells, adjust the metabolic function of body cells, enhance body immunity, and delay aging, etc. The human-derived stem cell bioactive substance group composition in the invention boasts substantial application prospects in medicine, healthcare and beauty maintaining fields.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to extracting stem cells from umbilical cord tissue and umbilical cord blood of healthy people, cultivating and amplifying them in vitro, and then using the stem cells to prepare and apply bioactive substances. Background technique [0002] Stem cells are a kind 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 multi-directional differentiation potential, which can be obtained from ...

Claims

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Application Information

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IPC IPC(8): C12P1/00C12N5/074
Inventor 张正前
Owner 张正前
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