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Human pluripotent stem cell exosomes loaded with resveratrol, preparation method and application thereof

A technology of human pluripotent stem cells and resveratrol, applied in the direction of non-embryo pluripotent stem cells, artificially induced pluripotent cells, animal cells, etc., can solve the problems that need to be further confirmed, and the treatment potential of diseases and injuries has not been fully utilized , achieve good disease treatment effect, strong proliferation and expansion ability, and realize the effect of industrialized production

Active Publication Date: 2022-04-01
上海睿泰生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since milk exosomes contain a large number of non-human genes and proteins, their safety through intravenous systemic injection still needs to be further confirmed, and the focus of this study is mainly on the drug carrier function of exosomes. The therapeutic potential of the substances carried by exosomes to diseases and injuries has not been fully utilized

Method used

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  • Human pluripotent stem cell exosomes loaded with resveratrol, preparation method and application thereof
  • Human pluripotent stem cell exosomes loaded with resveratrol, preparation method and application thereof
  • Human pluripotent stem cell exosomes loaded with resveratrol, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Culture of human embryonic stem cells (ESCs) and extraction and identification of exosomes

[0082] A layer of embryonic stem cell Matrigel (ESC-Qualified BD Matrigel, BD Sparks, MD, USA), ESCs were moved into the dish, and mTeSR1 serum-free medium (StemCell Vancouver, BC, Canada), in an incubator (37°C, 5% CO 2 , saturated humidity) culture, and collect the culture medium changed every day. Filter the medium through a 0.22 micron pore size filter membrane and centrifuge at 10,000g at 4°C for 30 minutes to remove cell debris; use a 100KD molecular weight ultrafiltration tube and centrifuge (3500g, 15min) to intercept exosomes in the concentrated supernatant to obtain exosomes concentrate; transfer the concentrate to a 30% sucrose / heavy water density pad (1.210 g / cm 3 ), centrifuge at 100,000g at 4°C for 210 minutes, collect the 5ml sucrose / heavy water density pad at the bottom, add PBS to dilute, transfer to an ultrafiltration centrifuge tube with a molecular weigh...

Embodiment 2

[0087] Culture of human induced pluripotent stem cells (iPSCs) and extraction and identification of exosomes

[0088] A layer of embryonic stem cell Matrigel (ESC-Qualified BD Matrigel, BD Sparks, MD, USA), iPSCs were moved into the dish, and mTeSR1 serum-free medium (StemCell Vancouver, BC, Canada), in an incubator (37°C, 5% CO 2 , saturated humidity) culture, and collect the culture medium changed every day. Filter the medium through a 0.22 micron pore size filter membrane and centrifuge at 10,000g at 4°C for 30 minutes to remove cell debris; use a 100KD molecular weight ultrafiltration tube and centrifuge (3500g, 15min) to intercept exosomes in the concentrated supernatant to obtain exosomes concentrate; transfer the concentrate to a 30% sucrose / heavy water density pad (1.210 g / cm 3 ), centrifuge at 100,000g at 4°C for 210 minutes, collect the 5ml sucrose / heavy water density pad at the bottom, add PBS to dilute, transfer to an ultrafiltration centrifuge tube with a mol...

Embodiment 3

[0093] Human ESC-derived exosomes (ESC-Exos) encapsulated resveratrol (Res) by co-incubation method

[0094] ESC-Exos solution is from Example 1.

[0095] Quantitative standard curve determination of Res: Accurately weigh 1mg of Res powder and dissolve it in 1mL of methanol, then dilute the solution with methanol to prepare 50μg / mL, 20μg / mL, 10μg / mL Res in methanol standard solution, HPLC injection. The chromatographic conditions are as follows:

[0096] Chromatographic column: Zorbax Extend C-18, 150*4.6μm, 5–micro

[0097] Mobile phase: methanol: water = 95:5

[0098] Flow rate: 1mL / min

[0099] Column temperature: room temperature

[0100] Detection wavelength: 305nm

[0101] After obtaining the corresponding experimental results, the peak area (PA) of the chromatographic peak was plotted as a function of the Res concentration (C, μg / mL) (see attached image 3 ), obtain the quantitative standard curve of Res under this chromatographic separation condition as follows:...

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Abstract

The invention relates to human pluripotent stem cell exosomes loaded with resveratrol, a preparation method and application thereof. The present invention provides the application of human pluripotent stem cell exosomes and resveratrol-loaded human pluripotent stem cell exosomes in the following aspects: as an application for preparing drugs or cosmetics for preventing stretch marks, repairing stretch marks, or treating stretch marks, The application as the preparation of skin anti-aging medicine or cosmetics, the application of medicine or cosmetics for the treatment of skin scars, skin freckles, acne, acne, acne and the like. Compared with the prior art, the present invention uses exosomes derived from embryonic stem cells and induced human pluripotent stem cells as the drug carrier of resveratrol, which can greatly improve the biological effect of resveratrol, and combined with human pluripotent stem cells The function of exosomes can play a better role in the treatment of diseases.

Description

technical field [0001] The invention belongs to the technical fields of cell biology, molecular biology and drug research and development, and in particular relates to human pluripotent stem cell exosomes loaded with resveratrol and its preparation method and application. Background technique [0002] Exosomes are a type of extracellular vesicles secreted by cells with a diameter of about 30-150nm, which carry a variety of biologically active substances from donor cells, including DNA, RNA, protein, etc., into effector cells, effectively regulating the body's cell signal transduction guide. Studies have shown that exosomes secreted by stem cells can physiologically repair or remove damaged, diseased and aging cells in the body by delivering biologically active substances derived from stem cells to effector cells, and play anti-inflammatory, immune-regulating, and promote effector cell proliferation, migration and differentiation , Promote angiogenesis and other functions. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/545A61K31/05A61P17/18A61P17/10A61P17/02A61P17/00A61K8/99A61K8/34A61Q19/08A61Q19/00C12N5/074
CPCA61K35/545A61K31/05A61K8/99A61K8/347A61Q19/08A61Q19/00C12N5/0696A61K2300/00
Inventor 汪泱吴复跃何振东
Owner 上海睿泰生物科技股份有限公司
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