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rADSCs loaded gelatin microspheres and preparation method and application thereof

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

Active Publication Date: 2019-08-30
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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  • rADSCs loaded gelatin microspheres and preparation method and application thereof
  • rADSCs loaded gelatin microspheres and preparation method and application thereof
  • rADSCs loaded gelatin microspheres and preparation method and application thereof

Examples

Experimental program
Comparison scheme
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 rADSCs loaded gelatin microspheres and a preparation method and application thereof. The preparation process comprises the steps of using mixed liquor of fat stem cells and a gelatin aqueous solution as an inner phase, using an oil phase as an outer phase, and through a micro-fluidic device, preparing microdroplets; and then enabling gelatinized microdroplets to have a cross-linking reaction with a genipin solution so as to obtain the rADSCs loaded gelatin microspheres. According to the rADSCs loaded gelatin microspheres, live rADSCs is encapsulated in situ, besides, high activity of the rADSCs can be maintained, so that healing of wounds of diabetes and formation of wound blood vessels can be well promoted. Besides, the rADSCs loaded gelatin microspheres further have good degradation effects, and during wound healing, rADSCs loaded gelatin microspheres are just basically degraded completely. The rADSCs loaded gelatin microspheres concurrently have favorable wound healing effect, blood vessel formation effect and degradation effect, have tremendous application prospects in the respects of wound healing and blood vessel formation, and can be prepared into wound healing medicines and blood vessel formation medicines for application.

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 Applications(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
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