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Composition containing exogenous mitochondrion as active ingredient, application thereof and cell repairing method

A technology of mitochondria and composition, which is applied in the direction of pharmaceutical formulations, cosmetic preparations, skin care preparations, etc., can solve the problems of target cytotoxicity, mitochondrial damage, mitochondria and cell membrane rupture, etc., to improve wrinkles and skin aging, promote Increased, long-lasting direct protective effect

Active Publication Date: 2016-04-27
TAIWAN MITOCHONDRION APPLIED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although the above method can promote the delivery of mitochondria into cells, the carriers used, such as membrane-penetrating peptides and liposomes, may cause mitochondria and cell membrane rupture, resulting in mitochondrial damage and target cell toxicity.

Method used

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  • Composition containing exogenous mitochondrion as active ingredient, application thereof and cell repairing method
  • Composition containing exogenous mitochondrion as active ingredient, application thereof and cell repairing method
  • Composition containing exogenous mitochondrion as active ingredient, application thereof and cell repairing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Fluorescent calibration of mitochondria

[0051] Transfect red fluorescent protein DsRed or green fluorescent protein (green fluorescent protein) with mitochondrial signal peptide (mitochondria signal peptide) into baby hamster kidney fibroblast cells (babyhamster kidney fibroblast cells, BHK-21 cells, hereinafter referred to as BHK cells), through G418 antibiotics and flow Screened by a cell sorter to obtain RedM-BHK cells or GFP-BHK cells that can continuously express red fluorescent protein.

Embodiment 2

[0052] Example 2: Isolation of mitochondria from BHK cells

[0053] When the cell number of BHK cells was raised to 2×10 8 At this time, the cell culture dish was washed by adding SEH buffer (0.25M sucrose, 0.5mM EGTA and 3mM HEPES-NaOH, pH 7.2), and centrifuged at 1000xg for 3 minutes. After the supernatant was removed, 2 ml of Grind SEH buffer about 15 times in a Dounce homogenizer and operate on ice to reduce damage to cells and mitochondria. After grinding, the homogeneous solution was centrifuged at 1000xg for 15 minutes to remove the precipitate, and then centrifuged at 9000xg for 10 minutes. Finally, the precipitate was dissolved in 50 μL of SEH buffer, and an inhibitor of proteolytic enzyme was added. Store at ℃.

Embodiment 3

[0054] Example 3: Determining the way for mitochondria to enter cells

[0055] In this example, in order to trace the movement path of mitochondria into the cell, the relationship between the moving position of mitochondria and lysosomes will be observed at different times by adding exogenous mitochondria.

[0056] First, mitochondria were marked with red fluorescent protein DsRed, and BHK cells were transfected with green fluorescent lysosome dye (LysoTracker) to determine the location of lysosomes in the cells. Administer 5 μg of exogenous mitochondria marked with red fluorescent protein, and co-culture with BHK cells that have been treated with lysosome dye at room temperature. After culturing for one hour and four hours, observe the entry of exogenous mitochondria into BHK under a conjugate focal microscope The situation of the cell and its relative position with the lysosome, the results are as follows figure 1 and figure 2 shown.

[0057] Depend on figure 1 It can b...

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PUM

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Abstract

The invention discloses a composition containing exogenous mitochondrion as active ingredient, application thereof and a cell repairing method. The composition comprises exogenous mitochondria and at least one pharmaceutically acceptable carrier or carrier acceptable in beauty, and further comprises an adjuvant which is prepared from a group composed of serum, blood plasma, complement and combination of at least two of serum, blood plasma and complement. The exogenous mitochondria are obtained from cells by a centrifugal purification way. The composition is of high safety. The exogenous mitochondria are sent into cells completely as an effective dose of the composition is fed into an individuality, thereby achieving cell repairing and cell senescence controlling or preventing.

Description

technical field [0001] The present invention relates to methods for anti-aging and repairing damaged mitochondria, in particular to a composition with exogenous mitochondria as an active ingredient, its use and a method for repairing cells. Background technique [0002] Mitochondria are responsible for providing cellular energy and producing adenosine triphosphate (ATP). Mitochondria will undergo dynamic changes due to differences in intracellular energy requirements or cellular stress, and are not always in a single mitochondrial state. Specifically, when the energy demand of cells increases, mitochondria will continue to divide (fission) to rapidly produce adenosine triphosphate; on the contrary, when cells are starved, mitochondria will undergo fusion (fusion) to reduce energy production and Use to maintain the normal physiological function of cells. In addition, when mitochondria encounter damage such as decreased membrane potential and mitochondrial DNA mutation (mtDN...

Claims

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Application Information

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IPC IPC(8): A61K8/98A61K8/97A61Q19/08
Inventor 苏鸿麟曾学敏吴诗芳
Owner TAIWAN MITOCHONDRION APPLIED TECH
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