Adult stem cell and its culturing method and use

A technology for adult stem cells and cell culture, applied in the field of adult stem cells and their cultivation, can solve problems such as new adult stem cell culture methods and application reports that have not yet been seen

Inactive Publication Date: 2005-09-14
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0022] So far, there have been no new reports on new adult stem cells derived from blood and their culture methods and uses.

Method used

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  • Adult stem cell and its culturing method and use
  • Adult stem cell and its culturing method and use
  • Adult stem cell and its culturing method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0193] Embodiment 1, the formula of Potentculture culture reagent

[0194] The specific composition and content of 1000ml of Potentculture culture reagent are as follows:

[0195] ① Amino acids: 1682.9mg

[0196] The amino acid products described below are from Gibico Company of the United States without labeling their sources.

[0197] A. 8 kinds of essential amino acids 592.4mg

[0198] Including 94.4mg of L-isoleucine (product number 22405013, purity>99%), 94.4mg of L-leucine (product number 12424016, purity>99%), 131.6mg of L-lysine (product number 12407011, Purity>99%), 26.9mg of methionine (US Sigma Company, product number 9625, purity 98%), 59.4mg of L-phenylalanine (product number 12409017, purity>99%), 15.7mg of L-tryptophan (Product No. 12432019, purity>99%), 85.7 mg of L-threonine (Prod. No. 12410015, purity>99%) and 84.3 mg of L-valine (Prod. No. 12413019, purity>99%).

[0199] B. Non-essential amino acids 1090.5mg

[0200] Including 15mg of L-alanine (product...

Embodiment 2

[0219] The culture method of embodiment 2, PSC3445

[0220] (1) Sources of main reagents: newborn horse serum and calcium-magnesium-free Hank’s solution were purchased from GIBCO Company in the United States, fetal bovine serum was from Hangzhou Sijiqing Company, and lymphocyte separation solution was purchased from Shanghai Reagent No. 2 Factory.

[0221] ① Separation of cord blood mononuclear cells: collect 80ml of fresh cord blood from normal people (healthy parturients in obstetrics and gynecology) under sterile conditions, anticoagulate with heparin (50U / ml); process within 2 hours after cord blood collection: fresh cord blood 2500r min -1 Centrifuge for 15 minutes, take the buffy coat (white blood cell layer); then dilute it with calcium and magnesium-free Hank's solution containing 2% fetal bovine serum at a ratio of 1:3, superimpose on the upper layer of Ficoll-paque, 3000r min -1 After centrifugation for 15 min, the mononuclear cell layer was taken and washed with ca...

Embodiment 3

[0226] The directional induction of embodiment 3, PSC3445

[0227] After the PSC3445 of the present invention is cultured for 10-12 days, an appropriate inducer is added for induction. The specific induction process and results of PSC3445 induced by four inducers are as follows:

[0228] ①Targeted induction of lymphocytes

[0229] The umbilical cord blood-derived PSC3445 cultured for 10-12 days was replaced with α-MEM containing 10% fetal bovine serum and 300 U / ml IL-2 for induction, and a control group (without adding IL-2 and without changing culture Reagent two control groups). Rehydration was performed once every three days, and the IL-2 concentration was still 300 U / ml during rehydration. After 7-10 days of induction, PSC3445 becomes a suspension growth cell, and the suspension cell is CD3 by flow cytometry detection and morphological observation + CD4 + or CD3 + CD8 + of T lymphocytes.

[0230] ② Directed induction of macrophages

[0231] PSC3445 derived from um...

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Abstract

This invention relates to one kind of new adult stem cell and its cultivation method and application. The cell has high self duplication capability and multiple system differentiation capability. Its cell cultivation feature, growth feature, morphologic feature and biological feature are different from existing adult stem cell. It has been oriented inducting differenciated to T lymphocyte, macrophage, epidermic cell and nerve cell. First cultivation second paste method is used in cultivation. So mass amplification high purity cell can be got by this inveniton. Clinical application prospect of this cell is wide, and it can be used to human body repair and rebuild, form artificial tissue, anti-decline, ant-cancer, and cure low hematopiesis function and low immunologic function. It can be used as gene carrier to cure bone damage or hematopiesis disturbance, it is the one of the first plan to solve problem of difficult stem cell source.

Description

technical field [0001] The present invention relates to the technical field of medicine, in particular to a new cell and its culture method and use, more specifically to an adult stem cell and its culture method and use. Background technique [0002] Stem cells are undifferentiated cells with two basic characteristics, one is self-replicating ability, and the other is the ability to differentiate into more than one type of functional cells. According to the size of differentiation potential, stem cells are generally divided into three categories. The first type is totipotent stem cells, that is, within the first hour after fertilization, a fertilized egg can divide into totipotent cells with the same function, and any of these cells can develop into a fetus if placed in a woman's uterus; The second type is embryonic stem cells (Embryonic stem cells), which have extensive differentiation potential and can generate all tissue cells in the body except the placenta; the third t...

Claims

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Application Information

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IPC IPC(8): A61K35/545C12N5/074
Inventor 赵书平沈茜于丽华张军徐玉莲李闻捷张俊洁
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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