A Chinese herbal medicine, mushroom, natural product or nutrient composition for promoting proliferation of stem cells and applications thereof

Abstract

The application discloses a Chinese herbal medicine, mushroom, natural product or nutrient composition for promoting stem cell proliferation and application thereof, and relates to the technical field of biotechnology, wherein the composition comprises ingredients or extracts selected from the following group: (a) Chinese herbal medicine; (b) medicinal mushroom; (c) active substance of natural origin; (d) nutrient; and the Chinese herbal medicine is at least one of ginseng, rhizoma polygonati, ganoderma lucidum, panax notoginseng, astragalus, angelica sinensis or medlar, wherein the composition is prepared by selecting and combining Chinese herbal medicine, medicinal mushroom, active substance or nutrient of natural origin, and provides a safe and reliable solution with high biocompatibility, and potential risks possibly caused by using exogenous growth factors or gene regulation are effectively avoided. The solution can realize multi-target synergistic effect, significantly promotes the proliferation rate of stem cells and maintains high activity of the stem cells.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a combination of traditional Chinese medicine, mushrooms, natural substances or nutrients that promotes stem cell proliferation and their applications. Background Technology

[0002] Stem cells are special cells with self-renewal and multi-lineage differentiation potential, forming the core foundation for tissue repair, regenerative medicine, and disease intervention. Cell proliferation is a crucial process for stem cells to realize their biological functions, referring to their ability to increase their number through cell division. In vivo, adequate stem cell proliferation ensures tissue homeostasis and damage repair; in vitro, efficient and stable proliferation is a prerequisite for stem cell expansion, research, and clinical applications. However, regardless of the in vivo microenvironment or in vitro culture system, stem cells face common challenges such as limited proliferation efficiency and susceptibility to inactivation or functional decline during long-term culture.

[0003] Current mainstream technologies for promoting stem cell proliferation heavily rely on recombinant growth factors (such as bFGF and EGF) or chemically defined inducers. While effective, these methods have significant limitations: high cost, potential introduction of exogenous risks (such as immunogenicity and batch-to-batch variability), and the potential for abnormal cell states with long-term use (such as increased genomic instability). Therefore, developing a safe, economical, easily standardized alternative that can simultaneously promote proliferation and maintain stem cell function has become an urgent technological need in this field. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a combination of traditional Chinese medicine, mushrooms, natural substances, or nutrients that promotes stem cell proliferation, and its application, thereby solving the technical problems mentioned in the background section.

[0005] To achieve the above objectives, the present invention is implemented through the following technical solution: a composition of traditional Chinese medicine, mushrooms, natural substances or nutrients that promotes stem cell proliferation, the composition comprising an ingredient or extract thereof selected from the group consisting of: (a) traditional Chinese medicine; (b) medicinal mushrooms; (c) naturally derived active substances; (d) nutrients; wherein the traditional Chinese medicine is at least one of ginseng, polygonatum, ganoderma, notoginseng, astragalus, angelica or wolfberry; and / or the medicinal mushroom is at least one of ganoderma, cordyceps, hericium erinaceus, agaricus or turkey tail mushroom.

[0006] In some embodiments, the naturally derived active substance is at least one of polysaccharides, saponins, flavonoids, polyphenols, alkaloids, or amino acids.

[0007] In some embodiments, the nutrient is at least one of vitamins, minerals, amino acids, fatty acids, or coenzymes.

[0008] In some embodiments, the composition comprises at least two of the following herbal extracts selected from the group consisting of ginseng extract, polygonatum extract, ganoderma extract, notoginseng extract, astragalus extract, angelica extract, and wolfberry extract.

[0009] In some embodiments, the composition further comprises at least one of the following medicinal mushroom extracts selected from the group consisting of Ganoderma lucidum extract, Cordyceps sinensis extract, Hericium erinaceus extract, Agaricus blazei extract, and Trametes versicolor extract.

[0010] In some embodiments, the composition further comprises vitamin C and / or amino acids.

[0011] In some embodiments, the composition is one or more physical forms selected from extracts, concentrates, fermentation products, powders, or solutions.

[0012] In some embodiments, the composition is used in the preparation of an agent for promoting stem cell proliferation, wherein the stem cells are selected from embryonic stem cells, adult stem cells, or mesenchymal stem cells.

[0013] This invention provides a composition of traditional Chinese medicine, mushrooms, natural substances, or nutrients that promotes stem cell proliferation, and its application. By carefully selecting and combining naturally sourced traditional Chinese medicine, medicinal mushrooms, active substances, or nutrients, this invention provides a safe, reliable, and highly biocompatible solution, effectively avoiding the potential risks associated with using exogenous growth factors or gene regulation. This approach exhibits multi-target synergistic effects, significantly promoting stem cell proliferation and maintaining their high activity. More importantly, its long-term application helps maintain stem cell stemness (self-renewal capacity) and multi-lineage differentiation potential, effectively delaying functional cellular aging. Attached Figure Description

[0014] Figure 1 This is a flowchart illustrating a combination of traditional Chinese medicine, mushrooms, natural substances, or nutrients that promotes stem cell proliferation, as described in this invention, and its application. Detailed Implementation

[0015] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0016] Please see Figure 1The present invention provides a technical solution: a composition of traditional Chinese medicine, mushrooms, natural substances or nutrients that promotes stem cell proliferation, the composition comprising an ingredient or extract thereof selected from the group consisting of: (a) traditional Chinese medicine; (b) medicinal mushrooms; (c) naturally derived active substances; (d) nutrients; wherein the traditional Chinese medicine is at least one of ginseng, polygonatum, ganoderma, notoginseng, astragalus, angelica or wolfberry; and / or the medicinal mushroom is at least one of ganoderma, cordyceps, hericium erinaceus, agaricus or turkey tail.

[0017] In some embodiments, the naturally derived active substance is at least one of polysaccharides, saponins, flavonoids, polyphenols, alkaloids, or amino acids.

[0018] In some embodiments, the nutrient is at least one of vitamins, minerals, amino acids, fatty acids, or coenzymes.

[0019] In some embodiments, the composition comprises at least two of the following herbal extracts selected from the group consisting of ginseng extract, polygonatum extract, ganoderma extract, notoginseng extract, astragalus extract, angelica extract, and wolfberry extract.

[0020] In some embodiments, the composition further comprises at least one of the following medicinal mushroom extracts selected from the group consisting of Ganoderma lucidum extract, Cordyceps sinensis extract, Hericium erinaceus extract, Agaricus blazei extract, and Trametes versicolor extract.

[0021] In some embodiments, the composition further comprises vitamin C and / or amino acids.

[0022] In some embodiments, the composition is one or more physical forms selected from extracts, concentrates, fermentation products, powders, or solutions.

[0023] In some embodiments, the composition is used in the preparation of an agent for promoting stem cell proliferation, wherein the stem cells are selected from embryonic stem cells, adult stem cells, or mesenchymal stem cells.

[0024] Its detailed connection method is a well-known technology in this field. The following mainly introduces the working principle and process, and the specific work is as follows.

[0025] Example 1: Effect of the Composition on Mesenchymal Stem Cell Proliferation 1. Composition Preparation: Ginseng aqueous extract (containing ginsenosides) and Ganoderma lucidum polysaccharide extract (containing β-glucan) were prepared according to conventional extraction methods in the art and freeze-dried into powder. The ginseng extract powder, Ganoderma lucidum extract powder, and pharmaceutical-grade vitamin C powder were accurately weighed at a mass ratio of 2:1:0.1 and thoroughly mixed in a mixer under aseptic conditions to obtain a uniform composition powder A. This formulation is based on the theory of "tonifying qi and strengthening the foundation" in traditional Chinese medicine and the theory of antioxidant support in modern nutrition, aiming to synergistically promote cell metabolism and vitality. 2. Cell Culture and Treatment: Mesenchymal stem cells derived from bone marrow of healthy individuals were obtained from commercial sources and routinely passaged using basal medium (DMEM / F12) containing 10% fetal bovine serum and 1% double antibiotics. Cells in the logarithmic growth phase of passages 3-5 were harvested and cultured at a ratio of 2:1:0.1 per well. Cells were seeded at a density in 96-well plates. After cell attachment, the medium was replaced with complete medium (sterilized by 0.22 μm filter) containing different concentrations of composition powder A. Experimental groups (containing a final composition concentration of 10 μg / mL, 50 μg / mL, and 100 μg / mL) and a control group (containing only an equal volume of solvent or blank medium) were set up. Each group had 6 replicates. 3. Proliferation assay: After culturing the cells in a 37°C, 5% CO2 incubator for 72 hours, cell proliferation was assessed using the widely accepted CCK-8 assay. Specifically, 10 μL of CCK-8 reagent was added to each well, and incubation was continued for 2 hours. The absorbance of each well was then measured at 450 nm using a microplate reader. The absorbance value was directly proportional to the number of viable cells. 4. Results analysis: The experimental data showed that compared with the control group, the absorbance values ​​of all experimental groups with added composition powder A were significantly increased (P<0.05), exhibiting a dose-dependent promoting effect. At a concentration of 100 μg / mL, the cell proliferation rate increased by approximately 35%-45% compared to the control group. The results indicate that the ginseng-Ganoderma lucidum-vitamin C composition described in this invention can effectively and safely promote the in vitro proliferation of mesenchymal stem cells.

[0026] Example 2: Evaluation of the Composition on Stem Cell Activity and Function Maintenance 1. Composition Preparation: Polysaccharide extract from Polygonatum sibiricum and β-glucan extract from Hericium erinaceus were prepared using a combination of ethanol extraction and hot water extraction. These two extracts were mixed with lentinan purified from natural plants at a mass ratio of 1.5:1:0.5 to prepare composition solution B (dissolved in PBS and filtered for sterilization). 2. Long-Term Culture Experiment: Human adipose-derived mesenchymal stem cells were continuously passaged under standard culture conditions. Starting from the 5th generation, the experimental group received 50 μg / mL of composition solution B in the culture medium, while the control group received no addition. Cells were passaged at each generation, and the population doubling time was recorded. Stem cell markers (such as the positive rates of CD73, CD90, and CD105, and the negative rates of CD34 and CD45) were periodically detected using flow cytometry. 3. Functional Detection: At the 10th generation, osteogenic and adipogenic differentiation induction experiments were performed on both groups of cells. After 21 days of induction, the multi-lineage differentiation potential was assessed by Alizarin Red S staining (to detect calcium nodules) and Oil Red O staining (to detect lipid droplets), respectively. 4. Results Analysis: Long-term culture results showed that the cell population doubling time in the experimental group supplemented with composition solution B was stable and shorter than that in the control group, indicating that the proliferation capacity was well maintained. Flow cytometry showed that the positive rate of stem cell markers in the experimental group remained at a high level (>95%), significantly better than that in the control group at the same time point (the positive rate decreased to about 80%). Differentiation experiments showed that the cells in the experimental group still retained good osteogenic and adipogenic differentiation capacity, and the staining positive rate was not significantly different from that of early passage cells, while the differentiation efficiency of the control group cells decreased significantly. This example demonstrates that the composition of the present invention can not only promote proliferation, but also help maintain the stem characteristics and multi-lineage differentiation function of stem cells in long-term culture, and delay their aging or functional decline.

[0027] Example 3: Application of Different Dosage Forms of the Composition 1. Dosage Form Preparation: The composition powder A from Example 1 was mixed with pharmaceutically acceptable excipients (such as microcrystalline cellulose and lactose) at a ratio of 1:9 and filled into capsule shells to prepare capsule C containing 100 mg of the composition per capsule. The composition powder A was also dissolved in sterile physiological saline to prepare injection solution D (terminally sterilized) with a concentration of 10 mg / mL. 2. In Vitro Simulation Application: For capsule C, the dissolution process in vivo after oral administration was simulated using pH gradient buffers to confirm that its main active ingredient could be effectively released in a simulated gastrointestinal environment. For injection solution D, proliferation experiments were conducted by directly adding it to stem cell culture medium at different dilutions (method as in Example 1). 3. Results and Explanation: The dissolution test showed that the release of the active ingredient from capsule C met expectations. In vitro cell experiments confirmed that appropriately diluted injection solution D culture medium also significantly promoted stem cell proliferation. This embodiment illustrates that the composition of the present invention can be prepared into different dosage forms (such as oral solid dosage forms and injections), and is expected to promote stem cell proliferation and maintain activity through corresponding administration routes (such as oral ingestion followed by in vivo metabolism and absorption to act on endogenous stem cells, or as a culture additive, or for local / systemic injection). This demonstrates its diverse application potential in the development of health foods, cell culture additives, or pharmaceutical preparations.

[0028] It should be noted that in this paper, relational terms such as first and second are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations.

Claims

1. A composition of traditional Chinese medicine, fungi, natural substances, or nutrients that promotes stem cell proliferation, characterized in that, The composition comprises an ingredient or extract thereof selected from the group consisting of: (a) traditional Chinese medicine; (b) medicinal fungi; (c) naturally derived active substances; and (d) nutrients; wherein the traditional Chinese medicine is at least one of ginseng, polygonatum, ganoderma, notoginseng, astragalus, angelica or wolfberry; and the medicinal fungi is at least one of ganoderma, cordyceps, hericium erinaceus, agaricus blazei or trachomatis.

2. The composition of traditional Chinese medicine, fungi, natural substances, or nutrients for promoting stem cell proliferation according to claim 1, characterized in that... The naturally derived active substance is at least one of polysaccharides, saponins, flavonoids, polyphenols, alkaloids, or amino acids.

3. The composition of traditional Chinese medicine, fungi, natural substances, or nutrients for promoting stem cell proliferation according to claim 1, characterized in that... The nutrients mentioned are at least one of vitamins, minerals, amino acids, fatty acids, or coenzymes.

4. A composition of traditional Chinese medicine, fungi, natural substances, or nutrients for promoting stem cell proliferation according to any one of claims 1-3, characterized in that... The composition comprises at least two of the following herbal extracts selected from the group consisting of ginseng extract, polygonatum extract, ganoderma extract, notoginseng extract, astragalus extract, angelica extract, and wolfberry extract.

5. A composition of traditional Chinese medicine, fungi, natural substances, or nutrients for promoting stem cell proliferation according to claim 4, characterized in that... The composition further comprises at least one of the following medicinal mushroom extracts selected from the group consisting of Ganoderma lucidum extract, Cordyceps sinensis extract, Hericium erinaceus extract, Agaricus blazei extract, and Trametes versicolor extract.

6. A composition of traditional Chinese medicine, fungi, natural substances, or nutrients for promoting stem cell proliferation according to claim 5, characterized in that... The composition also contains vitamin C and / or amino acids.

7. A combination of traditional Chinese medicine, fungi, natural substances, or nutrients for promoting stem cell proliferation according to any one of claims 1-6, and its application, characterized in that... The composition is one or more physical forms selected from extracts, concentrates, fermentation products, powders or solutions.

8. The application of a traditional Chinese medicine, fungus, natural substance, or nutrient composition for promoting stem cell proliferation according to any one of claims 1-7, characterized in that... The use of the composition in the preparation of an agent for promoting stem cell proliferation, wherein the stem cells are selected from embryonic stem cells, adult stem cells or mesenchymal stem cells.

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