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

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...

Examples

Embodiment 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% double antibody), seal it well, and refrigerate and transport it to the laboratory within 4 hours;

[0056] (2) In the clean room of the laboratory, check whether the aseptic bottle holding the umbilical cord tissue is complete, whether the transportation temperature is qualified, record the container label, delivery 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 aseptic bottle and wash it with normal saline for 5 times, keep the 1ml mark of the last washing solution and send it to the quality inspection room for bacteria inspection;

[0058] (4) Use ...

Embodiment 2

[0062] Embodiment 2: collection, centrifugation, ultrafiltration concentration of cell culture fluid

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

[0064] (2) Collect the centrifuged cell supernatant, first perform ultrafiltration dialysis through a 50KD ultrafiltration membrane, and take the dialysate; then pass the dialysate through a 5KD ultrafiltration membrane to collect the intercepted liquid, and the obtained liquid contains a large amount of cytokines of concentrate.

Embodiment 3

[0065] Example 3: Preparation of umbilical cord mesenchymal stem cell factor freeze-dried powder

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

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

[0068] (3) After the liquid is cooled to room temperature, add umbilical cord mesenchymal stem cell factor concentrate, and finally dilute to 1L with purified water, and stir until it becomes 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 packaging: the filtered liquid is divided into 2mL per vial (7mL) and half-filled with rubber stoppers (the containers used for the packaging must pass through 121°C, 20min, 0.1MPa Sterilized under...

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...

Examples

Embodiment 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% double antibody), seal it well, and refrigerate and transport it to the laboratory within 4 hours;

[0056] (2) In the clean room of the laboratory, check whether the aseptic bottle holding the umbilical cord tissue is complete, whether the transportation temperature is qualified, record the container label, delivery 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 aseptic bottle and wash it with normal saline for 5 times, keep the 1ml mark of the last washing solution and send it to the quality inspection room for bacteria inspection;

[0058] (4) Use ...

Embodiment 2

[0062] Embodiment 2: collection, centrifugation, ultrafiltration concentration of cell culture fluid

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

[0064] (2) Collect the centrifuged cell supernatant, first perform ultrafiltration dialysis through a 50KD ultrafiltration membrane, and take the dialysate; then pass the dialysate through a 5KD ultrafiltration membrane to collect the intercepted liquid, and the obtained liquid contains a large amount of cytokines of concentrate.

Embodiment 3

[0065] Example 3: Preparation of umbilical cord mesenchymal stem cell factor freeze-dried powder

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

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

[0068] (3) After the liquid is cooled to room temperature, add umbilical cord mesenchymal stem cell factor concentrate, and finally dilute to 1L with purified water, and stir until it becomes 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 packaging: the filtered liquid is divided into 2mL per vial (7mL) and half-filled with rubber stoppers (the containers used for the packaging must pass through 121°C, 20min, 0.1MPa Sterilized under...

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