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Stem cell-based medicinal product for treating diabetes and preparing method thereof

A technology of stem cell preparation and diabetes, which is applied in the field of stem cell research, can solve problems such as limited sources, and achieve the effects of abundant sources, convenient collection, and broad application prospects

Inactive Publication Date: 2016-04-06
SHENZHEN BEIKE BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the shortcomings of existing methods for treating diabetes, as well as the limited source and ethical issues involved in the process of obtaining stem cells, the present invention provides a stem cell preparation for treating diabetes and its preparation method

Method used

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  • Stem cell-based medicinal product for treating diabetes and preparing method thereof
  • Stem cell-based medicinal product for treating diabetes and preparing method thereof
  • Stem cell-based medicinal product for treating diabetes and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Preparation of mesenchymal stem cell preparations derived from umbilical cord

[0032] Operate in the ultra-clean workbench, sterilize the freshly obtained umbilical cord with ethanol, then wash the umbilical cord with saline, separate and strip the blood vessels of the umbilical cord to obtain the Wharton's glue part, cut the Wahton's glue into pieces, and dilute it into a bottle. Use the tissue adherence method to culture in a carbon dioxide incubator, observe the cell growth under a microscope, you can see that the cells grow adherently, are long fusiform, the peritoneum is transparent, the cell body is small, and has a three-dimensional effect, and it is gradually removed by changing the whole volume For non-adherent cells, the primary cells can be harvested when the cell confluence reaches 70-80%. The results are figure 1 ; The obtained primary cells were subcultured in DMEM medium to achieve the purpose of purifying and expanding mesenchymal stem cells. The results ar...

Embodiment 2

[0034] Study on Surface Markers of Mesenchymal Stem Cells Derived from Umbilical Cord

[0035] Use flow cytometry to detect cell surface markers, see the results image 3 . The cell surface markers CD73, CD90, and CD105 related to MSCs are highly expressed, and the expression of each generation is stable, all higher than 95%; cell surface markers CD14, CD34, CD45, CD19 related to hematopoietic cells and related to immune rejection The expression of HLA-DR is low, below 3%. It is proved that MSCs derived from umbilical cord after multiple passages contain stem cells that can proliferate stably. The cells highly express MSCs-related markers, but do not express or low-express hematopoietic cells and cell surface markers related to transplant rejection, suggesting that such MSCs cells are a type of low Immunogenic stem cells may play a role in fighting autoimmune diseases and treating rejection after transplantation.

Embodiment 3

[0037] Proliferation of mesenchymal stem cells derived from umbilical cord

[0038] The passage cells were planted in a 96-well plate, divided into 12 groups, each with 7 wells, and 200 μl of cell suspension was added to each well. Randomly take subcultured cells, detect the OD value of each experimental group and blank control group according to the MTT microplate reader, take the average, and draw the stem cell growth curve. The results are shown in Figure 4 . Experimental results show that mesenchymal stem cells can proliferate normally.

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Abstract

The invention discloses a stem cell-based medicinal product for treating diabetes. The stem cell-based medicinal product contains mesenchymal stem cells separated and extracted from the human umbilical cord, the expression of stem cell surface markers CD73, CD90 and CD105 is higher than 95%, and the expression of cell surface markers CD14, CD34, CD45, CD19 and HLA-DR is lower than 3%. The stem cell-based medicinal product can be differentiated into pancreatic beta cells under induction, so that insulin is secreted and blood sugar concentration is effectively reduced, and then a new method is provided for overcoming the defect that diabetes treatment depends on insulin and radically curing diabetes. The invention further provides a method for preparing the stem cell-based medicinal product. The method is easy to operate, raw material sources are wide, ethic limitation is avoided, and large-scale production can be conducted.

Description

Technical field [0001] The invention belongs to the field of stem cell research, in particular to a stem cell preparation for treating diabetes and a preparation method thereof. Background technique [0002] Diabetes is a series of clinical syndromes caused by absolute or relative lack of insulin in the body, mainly including type I and type II diabetes. In recent years, the incidence of diabetes has been on the rise, and it is becoming younger. It is estimated that by 2025, 334 million people worldwide will suffer from diabetes. Diabetes has become a major disease threatening human health. Different types of diabetes can cause the beta cells in the pancreas to fail to produce enough insulin to lower the blood sugar concentration, leading to hyperglycemia. In the current treatment methods, although drug treatment can effectively lower blood sugar, it cannot effectively repair islet function; although insulin injection can control symptoms, delay and reduce the occurrence of com...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/44A61P3/10C12N5/0775
Inventor 胡祥李陶刘沐芸李婵丁长才
Owner SHENZHEN BEIKE BIOTECH
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