A kind of preparation method of stem cell factor oral medicine

A stem cell factor and mesenchymal stem cell technology, applied in the biological field, can solve the problems of insufficient cytokine secretion process and small scale of cultivation, etc., and achieve the effects of enhancing human immunity, strengthening physique, and increasing absorption

Active Publication Date: 2022-08-02
安徽科门生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The pharmacological effects of stem cell factors are obviously reliable, but the culture medium or cell lysate collected from mesenchymal stem cells in vitro has problems such as small scale of culture and insufficient efficiency in the process of inducing cytokine secretion.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The preparation method of a stem cell factor oral medicine proposed by the present invention comprises the following steps:

[0022] S1. Obtain the fetal bovine umbilical cord, wash it with PBS, remove the arteries and veins, cut the umbilical cord into tissue blocks with a diameter of about 1-2 mm with scissors, add 0.4% collagenase II to digest for 2 h, centrifuge at 2000 r / min for 10 min, collect the precipitate, and add it to the 2% trypsin was added to digest for 30 min, the digestion was terminated, centrifuged at 2000 r / min for 15 min, and the precipitate was collected; PBS was pipetted to form a suspension, filtered through a 200 μm filter, and centrifuged at 1800 r / min for 7 min to obtain bovine umbilical cord cells;

[0023] S2. Inoculate bovine umbilical cord cells into a T25 culture flask, coat the culture flask with fibronectin, add α-MEM medium containing 10% FBS, 100U / mL penicillin, and 100U / mL streptomycin, at 37°C , 5%CO 2 Culture in an incubator, and ...

Embodiment 2

[0029] Compared with Embodiment 1, the difference is only in that step S3 is different; the details are as follows:

[0030] S3. When the P3 generation bovine umbilical cord stem cells are cultured until the cell confluence reaches 80-90%, the supernatant is discarded, washed with PBS, and then cultured with a secretory-promoting medium at a seeding density of 1.5×10 5 / mL, the supernatant was collected after 48 hours of induction culture, filtered with a 0.1 μm hollow fiber microfiltration membrane, and the permeate was concentrated by an ultrafiltration membrane (molecular weight cut-off 5 kD) to collect the retentate;

[0031] The secretory-promoting medium includes basal medium ɑ-MEM, and glutamine, 10 ng / mL norepinephrine, 2 ng / mL transforming growth factor, 60 ng / mL glutamine added to the basal medium at a final concentration of 3 ng / mL. / mL of glucosamine, 10ng / mL of astragalus polysaccharide.

Embodiment 3

[0033] Compared with Embodiment 1, the difference is only in that step S3 is different; the details are as follows:

[0034] S3. When the P3 generation bovine umbilical cord stem cells are cultured until the cell confluence reaches 80-90%, the supernatant is discarded, washed with PBS, and then cultured with a secretory-promoting medium at a seeding density of 1.5×10 5 / mL, the supernatant was collected after 48 hours of induction culture, filtered with a 0.1 μm hollow fiber microfiltration membrane, and the permeate was concentrated by an ultrafiltration membrane (molecular weight cut-off 5 kD) to collect the retentate;

[0035] The secretory-promoting medium includes basal medium DMEM / F12, and glutamine, 5ng / mL norepinephrine, 4ng / mL transforming growth factor, 80ng / mL glutamine added to the basal medium at a final concentration of 6ng / mL. / mL of glucosamine, 5ng / mL of astragalus polysaccharide.

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Abstract

The invention discloses a preparation method of a stem cell factor oral medicine, which relates to the field of biotechnology and includes the following steps: obtaining primary bovine umbilical cord mesenchymal stem cells; subculturing the primary bovine umbilical cord mesenchymal stem cells; When the umbilical cord stem cells are cultured until the cell confluency reaches 80-90%, use the secretory-promoting medium for culturing, collect the supernatant after culturing for 48 hours, filter with a 0.1 μm filter membrane, and then concentrate by ultrafiltration to obtain a concentrate; The culture medium includes basal culture medium and glutamine, norepinephrine, transforming growth factor, glucosamine, and astragalus polysaccharide added to the basal culture medium; the concentrated solution is pre-frozen, and then freeze-dried at ultra-low temperature to obtain freeze-dried powder . The invention can effectively improve the expansion of umbilical cord mesenchymal stem cells, induce the secretion of stem cell factors, and the obtained medicine can regulate the body from the cellular level, comprehensively improve the body function, improve sub-health, and repair and strengthen the immune system.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method of a stem cell factor oral medicine. Background technique [0002] The umbilical cord is the connecting structure between mammals and the fetus. It is shaped like a rope, with a smooth and transparent surface. It contains connective tissue, an umbilical vein and a pair of umbilical arteries. In recent years, with the continuous development of research, it has been possible to extract stem cells from the umbilical cord, cord blood and placenta. [0003] Umbilical cord mesenchymal stem cells refer to a kind of pluripotent stem cells existing in umbilical cord tissue. Umbilical cord mesenchymal stem cells are a type of low-immunogenic cells with multi-directional differentiation potential and strong immune regulation functions. Simple. An important function of umbilical cord mesenchymal stem cells is to secrete a variety of cytokines and growth factors in paracrin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/28A61K9/19A61P37/04C12N5/0775
CPCA61K35/28A61K9/19A61P37/04C12N5/0665C12N2500/32C12N2501/81C12N2501/90C12N2501/148C12N2501/15C12N2500/34
Inventor 张正亮
Owner 安徽科门生物科技有限公司
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