Application of umbilical cord mesenchymal stem cells to preparation of stem cell preparation for treating corneal injury

A stem cell preparation and quality stem cell technology, applied in the application field of stem cell preparations, can solve the problems such as the cornea cannot be restored to transparency, the rejection is severe, and the curative effect is poor, and achieves effective repair of corneal epithelial tissue, weak immunogenicity, and recovery. The effect of integrity and transparency

Inactive Publication Date: 2017-01-04
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for patients with severe corneal alkali burns, autologous corneal transplantation often requires a larger graft piece, which is taken from the healthy side eye. How much corneal limbal tissue can be taken from the healthy eye to ensure that the ocular surface stability of the healthy eye is not affected and effective Treatment of affected eyes remains an unresolved problem
In addition, for those injured with extensive ocular surface damage in both eyes, there are no autologous limbal stem cells available for transplantation; ② allogeneic limbal stem cell transplantation: although sufficient corneal limbal stem cell transplants can be provided, the source is limited and the its application
It should not be neglected that because the donor corneal limbal stem cell transplantation of this method has abundant capillaries, the rejection reaction after transplantation is often severe due to the lack of immune pardon; This method is widely used, but for patients with severe corneal injury such as corneal alkali burns, because the transparency of the amniotic membrane is worse than that of the cornea, even if the transplantation is successful, the cornea cannot return to its original transparency, and the curative effect is still not good

Method used

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  • Application of umbilical cord mesenchymal stem cells to preparation of stem cell preparation for treating corneal injury
  • Application of umbilical cord mesenchymal stem cells to preparation of stem cell preparation for treating corneal injury
  • Application of umbilical cord mesenchymal stem cells to preparation of stem cell preparation for treating corneal injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Preparation of umbilical cord-derived mesenchymal stem cell preparation

[0038] Operate in an ultra-clean workbench, sterilize the freshly obtained umbilical cord with ethanol, wash the umbilical cord with normal saline, separate and strip off the umbilical cord blood vessels, obtain the Wharton's jelly part, cut the Wharton's jelly into pieces, dilute it, and plant it in a bottle. Cultured in a carbon dioxide incubator using the tissue-attachment method. Observe the growth of the cells under a microscope. It can be seen that the cells are attached to the wall and grow in the shape of a long spindle. The cytoplasm is clear, the cell body is small, and has a three-dimensional effect. For non-adherent cells, the primary cells can be harvested when the cell fusion degree reaches 70-80%; the obtained primary cells are subcultured in DMEM medium to achieve the purpose of purifying and expanding the culture of mesenchymal stem cells, the results See figure 1 .

Embodiment 2

[0039] Example 2: Research on surface markers of umbilical cord-derived mesenchymal stem cells

[0040] The surface markers of the second-generation umbilical cord mesenchymal stem cells obtained in Example 1 were detected by flow cytometry, and the results are shown in Table 1 and figure 2 . Cell surface markers CD73, CD90, and CD105 related to MSCs were highly expressed, and the expressions of each generation were stable, all higher than 95%; cell surface markers related to hematopoietic cells CD11b, CD34, CD45, CD19 and immune rejection related The expression of HLA-DR was low, all less than 3%. It has been proved that MSCs derived from umbilical cord after multiple passages contain stem cells that can stably proliferate. The cells highly express MSCs-related markers, and do not express or low-express hematopoietic cells and cell surface markers related to transplant rejection, suggesting that this type of MSCs is a kind of low-grade MSCs. Immunogenic stem cells.

[004...

Embodiment 3

[0043] Example 3: Study on proliferation of umbilical cord-derived mesenchymal stem cells

[0044] Take P5 generation UC-MSC and inoculate in 24-well plate, 1×10 4 cells / hole. 3 wells were randomly collected and counted every day for 7 consecutive days to make growth curves. The results are shown in Table 2 and image 3 . The results showed that the growth incubation period was 0-4 days after inoculation of the P5 generation cells; about 4-7 days, the cell growth was active, which was a logarithmic growth period; after that, the cell growth entered the plateau period, and the cell growth curve was S-shaped.

[0045] Table 2. UC-MSCs cell count results

[0046]

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Abstract

The invention relates to the technical field of stem cells, in particular to application of umbilical cord mesenchymal stem cells to preparation of a stem cell preparation for treating corneal injury. Studies show that the umbilical cord mesenchymal stem cells can promote the corneal epithelium healing; the cornea turbid degree is low; the generation of cornea blood vessels is reduced; the cornea epithelial tissue can be more effectively repaired; the umbilical cord mesenchymal stem cells can be used for treating cornea alkali burn. The stem cells in the preparation are from the umbilical cord; the collection is convenient and is noninvasive operation; the obtained mesenchymal stem cells are infantile than other mesenchymal stem cells; the multiplication and differentiative potential is great; the umbilical cord resource is rich, and the large-scale production can be realized; the ethic limitation is avoided; the immunogenicity is low; the wide application prospects are realized in the field of corneal injury treatment.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to the application of umbilical cord mesenchymal stem cells in the preparation of stem cell preparations for treating corneal injuries. Background technique [0002] Ocular surface damage is one of the main causes of blindness worldwide. Among them, the most common causes are ocular chemical burns (such as alkali and acid burns) and thermal burns. This type of damage to the ocular surface is serious and difficult to treat. , the prognosis is poor, often leading to blindness and even loss of eyeballs. [0003] Corneal alkali burns are the most serious among chemical burns. Alkaline substances can dissolve in both fat and water, and can quickly undergo saponification reaction with lipids in cells, and the compounds formed can quickly penetrate hydrophilic The corneal stroma can quickly penetrate the lipophilic conjunctiva, corneal epithelium and endothelium, leading to liquefactio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/51A61K9/00A61P27/02
CPCA61K9/0019A61K9/0051A61K35/28A61K35/51
Inventor 葛啸虎陈海佳王一飞陈婉玲张维敏
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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