Preparation of umbilical cord mesenchymal stem cell factors with hair growth effect

A technology of mesenchymal stem cells and umbilical cord, which is applied in the field of preparation of umbilical cord mesenchymal stem cell factors, can solve the problems of unguaranteed use safety, short storage time, and reduced activity, so as to avoid social and ethical disputes, simple acquisition, The effect of stable storage

Inactive Publication Date: 2020-01-17
陕西中鸿科瑞再生医学研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] Direct use of umbilical cord mesenchymal stem cell culture medium can reduce the loss of active ingredients during repeated operations. However, some of the cytokines secreted by stem cells will reduce or lose their activity at room temperature or under temperature changes, which cannot reach Expecte

Method used

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  • Preparation of umbilical cord mesenchymal stem cell factors with hair growth effect
  • Preparation of umbilical cord mesenchymal stem cell factors with hair growth effect
  • Preparation of umbilical cord mesenchymal stem cell factors with hair growth effect

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0053] Example 1:

[0054] Step 1: Primary isolation and culture of umbilical cord mesenchymal stem cells

[0055] (1) Put the qualified umbilical cord tissue in a sterile bottle, containing 200ml of tissue preservation solution (α-MEM medium containing 2% bi-antibody), seal it, and refrigerate it to the laboratory within 4 hours;

[0056] (2) In the clean space of the laboratory, check whether the sterile bottles containing umbilical cord tissue are complete, whether the transportation temperature is qualified, record the container label, transportation time, and temperature related information in detail, observe the color of the umbilical cord, measure the length of the umbilical cord, and take photos for storage;

[0057] (3) In the biological safety cabinet, put the umbilical cord into a new sterile bottle and wash it 5 times with normal saline, save 1ml of the last washing solution and send it to the quality inspection room for bacteria inspection;

[0058] (4) Use pointed straight...

Example Embodiment

[0062] Example 2: Collection, centrifugation, ultrafiltration and concentration of cell culture fluid

[0063] (1) Take the cells of P4 generation, wait for the cells to grow to 85%, collect the cell culture solution and centrifuge in a centrifuge bottle, 800rpm, 5min;

[0064] (2) Collect the cell supernatant after centrifugation, first pass through a 50KD ultrafiltration membrane for ultrafiltration and dialysis, and then take the dialysate; then pass the dialysate through a 5KD ultrafiltration membrane to collect the interception fluid, and the resulting liquid contains a lot of cytokines The concentrate.

Example Embodiment

[0065] Example 3: Preparation of lyophilized powder of umbilical cord mesenchymal stem cell factor

[0066] (1) Weigh 100g of mannitol, heat it to 60-70℃, and dissolve it until clear.

[0067] (2) Take another beaker, weigh 4g of dextran 40, dissolve dextran 40 with 10% volume of purified water, heat and boil until it is completely clear, mix dextran and mannitol, and then add 6g of trehalose , Stir evenly until clear.

[0068] (3) After the liquid is brought to room temperature, add the umbilical cord mesenchymal stem cell factor concentrate, and finally make the volume to 1L with purified water, and stir until it is clear.

[0069] (4) Liquid filtration: the uniformly dissolved liquid is filtered with a 0.22 μm filter, and the filtrate is collected in a sterilized container.

[0070] (5) Liquid dispensing: the filtered liquid is divided into 2mL for each vial of vial (7mL), and a rubber stopper is added halfway after dispensing (the utensils used for dispensing must pass 121℃, 20min,...

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Abstract

The invention relates to the field of cytokine germination, in particular to preparation of umbilical cord mesenchymal stem cell factors with hair growth effect. The preparation includes the followingsteps: primary isolation and culture of human umbilical cord mesenchymal stem cells, collection of a cell culture supernatant, centrifugation, ultrafiltration and concentration to obtain a liquid containing the umbilical cord mesenchymal stem cell factor complex, and a freeze-drying process to prepare umbilical cord mesenchymal stem cell factor lyophilized powder. By the preparation method of theumbilical cord mesenchymal stem cell factors, activity of the cell factors can be preserved to the maximum and the cell factors are convenient to store and use. Hair follicle cells are fundamentallyactivated, and the hair regeneration effect is promoted remarkably and is more durable.

Description

technical field [0001] The invention relates to the field of cytokine hair growth, in particular to the preparation of an umbilical cord mesenchymal stem cell factor with hair growth effect Background technique [0002] Mesenchymal stem cells (mesenchymal stem cells, MSCs) are derived from the mesoderm in early development, and are a type of cells with self-replication and multilineage differentiation capabilities. Mesenchymal stem cells widely exist in many tissues throughout the body. Bone marrow, fat, synovium, dental pulp, amniotic fluid, placenta, umbilical cord, embryo, etc. can be isolated to prepare mesenchymal stem cells. Mesenchymal stem cells from different sources have similar morphology, express the same surface markers, and have similar biological characteristics. In addition to the multi-directional tissue cell differentiation ability of MSC, it also plays an important regulatory role in important functions of the human body such as hematopoiesis, immune infl...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K9/19A61K38/18A61P17/14C12N5/0775
CPCA61K9/19A61K35/28A61K38/18A61P17/14C12N5/0668C12N2509/00
Inventor 王花明磊国董玲娟
Owner 陕西中鸿科瑞再生医学研究院有限公司
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