Stem cell target-locating and enriching method

Abstract

The stem cell target locating and enriching method includes the following steps; marking the stem cell with 30-70 nm size immunological magnet bead to obtain marked stem cell; injecting the marked stem cell into the circulation system of target tissue; and aligning the external part corresponding to the target tissue to two dislike magnetic poles so as to enrich the stem cell in the target tissue during the circulating process of the circulation system, with the magnetic field strength between two dislike magnetic poles being 9200-24500 Gs. The present invention utilizes the magnetic characteristic of superparamagnetic bead in locating and enriching the destination cell in target tissue by means of magnetic targeting technology, and has the advantages of high enriching efficiency, simple process, etc. and thus wide application foreground.

Description

technical field

[0001] The invention relates to a method for targeting and enriching stem cells. Background technique

[0002] With the development of organ transplantation, transplantation immunology and the development and application of immunosuppressants, organ transplantation has become an effective means of treating clinical terminal diseases. Orthotopic liver transplantation is currently the only effective treatment for end-stage liver failure caused by various reasons (including poisons, drug poisoning, genetic metabolic diseases, etc.). However, due to the extreme shortage of liver donors, the high mortality rate associated with surgery and transplantation itself, the life-long administration of immunosuppressants and the serious or even fatal complications, the widespread clinical application of liver transplantation is greatly limited.

[0003] With the deepening of stem cell biology research, cell therapy using stem cells as seed cells has been widely used in di...

Examples

Embodiment 1

[0015] Example 1, c-Met + beta 2 m - Targeted enrichment of cells

[0016] In this example, the liver of the rat model of liver injury is used as the target tissue, and other tissues may also be used as the target tissue.

[0017] The immunomagnetic beads used in this example were purchased from the School of Materials, Tianjin University. The magnetic beads were coupled with goat anti-rabbit IgG, with a particle size of about 50 nm and good superparamagnetism.

[0018] 1. Obtaining rat bone marrow mononuclear cells

[0019] Rats sacrificed by neck dissection were sterilized with 75% ethanol, and the femur was taken out by laparotomy. The DMEM culture solution containing 10% fetal bovine serum was used as the flushing solution, and the bone marrow cavity was washed repeatedly with a 5ml syringe, and filtered through a 100-mesh sieve. The obtained cells were made into rat bone marrow single cell suspension, and after centrifugation at 1000rpm, the cells were resuspended in ...

Embodiment 2

[0029] Embodiment 2, detection of cell enrichment effect

[0030] 1. In vitro enrichment

[0031] According to the operation of steps 1-2 in Example 1, the immunomagnetic bead-labeled β 2 m -Cells were washed with PBS. Physiological saline containing 20% ​​dextran was then prepared as a mobile phase, and the resulting cells were suspended therein. Adjust the flow rate of the peristaltic pump to 0.1-15cm 3 / s, let the cells labeled with magnetic beads flow through the magnetic field (adjust the distance between the two magnetic poles to 1.6-2.5cm, so that the field strength fluctuates at 1.24-0.98T), count the ratio of the outflow and inflow cells under the microscope, and obtain the magnetic field to the magnetic bead Enrichment efficiency of labeled cells. The operation process is shown in Figure 4, and 15 in the figure is the β with immunomagnetic beads labeling 2 m + 20% dextran physiological saline solution for cells; 16 is a peristaltic pump; 17 is two magnetic pol...

technical field

[0001] The invention relates to a method for targeting and enriching stem cells. Background technique

[0002] With the development of organ transplantation, transplantation immunology and the development and application of immunosuppressants, organ transplantation has become an effective means of treating clinical terminal diseases. Orthotopic liver transplantation is currently the only effective treatment for end-stage liver failure caused by various reasons (including poisons, drug poisoning, genetic metabolic diseases, etc.). However, due to the extreme shortage of liver donors, the high mortality rate associated with surgery and transplantation itself, the life-long administration of immunosuppressants and the serious or even fatal complications, the widespread clinical application of liver transplantation is greatly limited.

[0003] With the deepening of stem cell biology research, cell therapy using stem cells as seed cells has been widely used in di...

Examples

Embodiment 1

[0015] Example 1, c-Met + beta 2 m - Targeted enrichment of cells

[0016] In this example, the liver of the rat model of liver injury is used as the target tissue, and other tissues may also be used as the target tissue.

[0017] The immunomagnetic beads used in this example were purchased from the School of Materials, Tianjin University. The magnetic beads were coupled with goat anti-rabbit IgG, with a particle size of about 50 nm and good superparamagnetism.

[0018] 1. Obtaining rat bone marrow mononuclear cells

[0019] Rats sacrificed by neck dissection were sterilized with 75% ethanol, and the femur was taken out by laparotomy. The DMEM culture solution containing 10% fetal bovine serum was used as the flushing solution, and the bone marrow cavity was washed repeatedly with a 5ml syringe, and filtered through a 100-mesh sieve. The obtained cells were made into rat bone marrow single cell suspension, and after centrifugation at 1000rpm, the cells were resuspended in ...

Embodiment 2

[0029] Embodiment 2, detection of cell enrichment effect

[0030] 1. In vitro enrichment

[0031] According to the operation of steps 1-2 in Example 1, the immunomagnetic bead-labeled β 2 m -Cells were washed with PBS. Physiological saline containing 20% ​​dextran was then prepared as a mobile phase, and the resulting cells were suspended therein. Adjust the flow rate of the peristaltic pump to 0.1-15cm 3 / s, let the cells labeled with magnetic beads flow through the magnetic field (adjust the distance between the two magnetic poles to 1.6-2.5cm, so that the field strength fluctuates at 1.24-0.98T), count the ratio of the outflow and inflow cells under the microscope, and obtain the magnetic field to the magnetic bead Enrichment efficiency of labeled cells. The operation process is shown in Figure 4, and 15 in the figure is the β with immunomagnetic beads labeling 2 m + 20% dextran physiological saline solution for cells; 16 is a peristaltic pump; 17 is two magnetic pol...

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