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A kind of gelatin microspheres loaded with radscs and preparation method and application thereof

A gelatin and microsphere technology, applied in the field of interventional medicine, can solve the problems of lack and limited application, and achieve the effects of good degradation, high activity and good wound healing.

Active Publication Date: 2021-08-13
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a kind of gelatin microspheres loaded with rADSCs in view of the lack of suitable cell carriers to load rADSCs in the prior art, which leads to the limitation of its application

Method used

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  • A kind of gelatin microspheres loaded with radscs and preparation method and application thereof
  • A kind of gelatin microspheres loaded with radscs and preparation method and application thereof
  • A kind of gelatin microspheres loaded with radscs and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0035] The preparation of embodiment 1 gelatin microsphere

[0036] 1. Fabrication of microfluidic devices

[0037] Prepare a PTFE tube with a length of about 35cm (inner diameter 0.3mm, outer diameter 0.6mm), insert a 26G needle into one end of the PTFE tube, insert a 31G needle into the PTFE tube in parallel at a distance of 20cm from the needle, and keep the needle tip in the middle of the tube; use AB Fix the needle and PTFE tube at the joint with rubber seal. After the device is completed, first check the connectivity and tightness of the device. Use a 5mL syringe to absorb deionized water, insert a 31G needle, and manually push the syringe. If liquid flows out of the PTFE tube, then Internal connectivity is good. Connect the syringes to the 20G needle at the same time, block the outlet of the PTFE tube, and squeeze the two syringes manually at the same time. If no liquid leaks out, the device is well sealed. Dry the prepared device for later use.

[0038] 2. Preparati...

Embodiment 2

[0047] Example 2 Preparation of gelatin microspheres loaded with rADSCs

[0048] The preparation of gelatin microspheres loaded with rADSCs, its specific preparation process is:

[0049] S1. Preparation of internal phase: rADSCs (rat adipose stem cells) were amplified in vitro to the P3-P4 generation, trypsinized the adherent cells in the culture flask, centrifuged after counting, added medium and resuspended to 10 7 cells / mL; then configure 20% gelatin solution, mix the two in the same volume, gently pipette and mix evenly, this is the internal phase, use a 1mL syringe to draw 100 μL of the internal phase solution each time, to prevent cells from being in the syringe during the preparation of microspheres Sinking leads to uneven dispersion;

[0050] The outer phase is peanut oil; the collection phase: add 1mL peanut oil to a 5mL round bottom EP tube, and pre-cool in ice water at 8°C;

[0051] Connect the microfluidic device, adjust the parameters of the syringe pump, contro...

Embodiment 3

[0053] Example 3 Detection of stem cell activity in gelatin microspheres loaded with rADSCs

[0054] The rADSC / MS prepared in Example 2 was added to the medium and cultured in a 37°C incubator. On the first and seventh days of culture, the Calcein-AM / PI staining solution was used for dead and live staining, and the cell viability was observed after taking pictures with CLSM.

[0055] The cell activity results of rADSC / MS on the 7th day of in vitro culture are as follows: figure 2 shown. From figure 2 It can be seen that after 7 days of in vitro culture and staining with Calcein-AM / PI, the microspheres were scanned by CLSM for 100 layers and then three-dimensionally reconstructed. Microspheres can effectively maintain the activity of adipose stem cells.

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Abstract

The invention discloses a gelatin microsphere loaded with rADSCs, a preparation method and application thereof. The preparation process is as follows: using the mixture of adipose stem cells and gelatin aqueous solution as the internal phase; using the oil phase as the external phase, micro-droplets are prepared through a microfluidic device; and then the micro-droplets are gelled and then cross-linked with the genipin solution After the reaction, gelatin microspheres loaded with rADSCs can be obtained. The gelatin microspheres loaded with rADSCs of the present invention can encapsulate live rADSCs in situ, and can maintain high activity of rADSCs at the same time, so that it can well promote the healing of diabetic wounds and the formation of wound blood vessels, and also has a good degradation effect , during wound healing, the gelatin microspheres loaded with rADSCs also happened to degrade almost completely. The gelatin microspheres loaded with rADSCs of the present invention have both good wound healing, angiogenesis and degradation effects, have great application prospects in wound healing and angiogenesis, and can be prepared as wound healing drugs and angiogenesis drugs to apply.

Description

technical field [0001] The invention relates to the technical field of interventional medicine, and more specifically relates to a gelatin microsphere loaded with rADSCs and a preparation method and application thereof. Background technique [0002] Transplantation of mesenchymal stem cells (MSCs) has emerged as a promising therapeutic strategy. MSCs can be stably expanded ex vivo while maintaining the ability to differentiate into epidermal and dermal lineage cells, and even skin appendages such as hair follicles, sweat glands, and microvessels can be rapidly regenerated in MSC-treated wounds. Injecting the cells directly into the repair site minimizes the invasiveness of the procedure. However, in this way, the degree of integration of cells into host tissues is low, and the cell survival rate is low, resulting in a low success rate of clinical transplantation. [0003] To address the above issues, a potentially attractive strategy is to suspend stem cells in hydrogels, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/16A61K47/42A61K35/28A61P17/02A61P3/10
CPCA61K9/0014A61K9/1658A61K35/28A61P3/10A61P17/02
Inventor 李燕史明张浩宋婷高芸芬
Owner SUN YAT SEN UNIV