Preparation method of stem cell factor sustained release microspheres for repairing endometrial injury

A technology of stem cell factor and slow-release microspheres, which is applied in the field of preparation of stem cell factor slow-release microspheres, can solve problems such as difficult to prevent adhesions and unstable repair of endometrial damage, and achieve uniform factor coating and prolong tissue Good separation effect and biocompatibility

Active Publication Date: 2022-05-13
广州喜舍俪健康管理咨询有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the direct use of exosome solution or factor solution for endometrial repair has the problem of sudden release, which cannot stably repair endometrial damage, and at the same time, the factor in the form of solution or exosome itself is difficult to play an anti-adhesion effect.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 1. Culture of placental mesenchymal stem cells:

[0021] 1. Under sterile conditions, take the fetal surface tissue of the full-term normal placenta, and wash it repeatedly with PBS until the liquid is clear.

[0022] 2. Ophthalmic scissors cut the placental tissue into pieces of about 1mm*1mm*1mm.

[0023] 3. Then use 0.1% type II collagenase to immerse the tissue fragments and digest them in a constant temperature shaking box at 37°C for 45 minutes.

[0024] 4. Grind the digested placental tissue through a 200-mesh sieve to collect the cell suspension, centrifuge at 1000 rpm / min at room temperature for 5 minutes, and discard the supernatant.

[0025] 5. Resuspend the cells with 2ml of phosphate buffered saline, add an equal amount of human lymphocyte separation medium, centrifuge at 2000rpm / min for 10 minutes, and absorb the buffy coat layer as much as possible.

[0026] 6. Use serum-free culture medium to adjust the cell concentration to 1*10 8 L -1 , inoculated ...

Embodiment 2

[0041] The difference from Example 1 is:

[0042] 1. Culture of placental mesenchymal stem cells:

[0043] 1. Under sterile conditions, take the fetal surface tissue of the full-term normal placenta, and wash it repeatedly with PBS until the liquid is clear.

[0044] 2. Ophthalmic scissors cut the placental tissue into pieces of about 1mm*1mm*1mm.

[0045] 3. Then use 0.1% type II collagenase to immerse the tissue fragments and digest them in a constant temperature shaking box at 37°C for 45 minutes.

[0046] 4. Grind the digested placental tissue through a 200-mesh sieve to collect the cell suspension, centrifuge at 1000 rpm / min at room temperature for 5 minutes, and discard the supernatant.

[0047] 5. Resuspend the cells with 2ml of phosphate buffered saline, add an equal amount of human lymphocyte separation medium, centrifuge at 2000rpm / min for 10 minutes, and absorb the buffy coat layer as much as possible.

[0048] 6. Use serum-free culture medium to adjust the cell ...

Embodiment 3

[0063] 1. Culture of placental mesenchymal stem cells:

[0064] 1. Under sterile conditions, take the fetal surface tissue of the full-term normal placenta, and wash it repeatedly with PBS until the liquid is clear.

[0065] 2. Ophthalmic scissors cut the placental tissue into pieces of about 1mm*1mm*1mm.

[0066] 3. Then use 0.1% type II collagenase to immerse the tissue fragments and digest them in a constant temperature shaking box at 37°C for 45 minutes.

[0067] 4. Grind the digested placental tissue through a 200-mesh sieve to collect the cell suspension, centrifuge at 1000 rpm / min at room temperature for 5 minutes, and discard the supernatant.

[0068] 5. Resuspend the cells with 2ml of phosphate buffered saline, add an equal amount of human lymphocyte separation medium, centrifuge at 2000rpm / min for 10 minutes, and absorb the buffy coat layer as much as possible.

[0069] 6. Use serum-free culture medium to adjust the cell concentration to 1*10 8 L -1 , inoculated ...

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PUM

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Abstract

The invention discloses a preparation method of stem cell factor sustained-release microspheres for repairing endometrial injury, which comprises the following steps: mixing a mesenchymal stem cell culture solution, sodium alginate and polyethylene glycol 4000 to obtain a water-phase solution, then mixing the water-phase solution with an emulsifier, and preparing the stem cell factor sustained-release microspheres under a stirring condition. According to the invention, sodium alginate and PEG4000 are used as materials to prepare the microspheres wrapping the stem cell factors. The adopted sodium alginate raw material is a natural polymer material, is good in biocompatibility, convenient to obtain and non-animal-derived, and is relatively slow to degrade in vivo compared with hyaluronic acid, gelatin and other raw materials. The PEG4000 can endow the microspheres with high viscoelasticity, can be quickly adhered to tissues, can cover intimal tissues, and can prevent the damaged endometrium from being contacted and adhered to the surroundings. In addition, the PEG4000 can delay absorption and prolong the tissue separation effect by gathering the protein of the intrauterine membrane, so that the auxiliary anti-adhesion effect is achieved.

Description

technical field [0001] The invention relates to stem cell factor slow-release microspheres, in particular to a preparation method of stem cell factor slow-release microspheres for repairing endometrial damage. Background technique [0002] In women, various pathological factors such as uterine cavity manipulation, uterine ischemia, and infection can cause endometrial damage. Endometrial repair impairment is an important cause of intrauterine adhesions, amenorrhea, and infertility in women. Currently, there are many methods to promote endometrial injury repair, but clinical treatment methods for severe endometrial injury repair impairment are also limited. The endometrium is a layer of mucous membrane that wraps the uterine cavity and is mainly divided into a functional layer and a basal layer. The proliferation and disintegration of the endometrium is regulated by the menstrual cycle. Decreased expression of estrogen and progesterone triggers endometrial shedding and bleed...

Claims

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Application Information

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IPC IPC(8): A61K9/52A61K35/28A61K47/36A61K47/10A61P15/00
CPCA61K9/5089A61K9/5036A61K9/5031A61K35/28A61P15/00
Inventor 蔡进辉
Owner 广州喜舍俪健康管理咨询有限公司
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