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Stem cell preparation for treating diabetes mellitus and preparation method of stem cell preparation

A stem cell preparation, stem cell technology, applied in the field of stem cell research to achieve the effect of not being ethically restricted, avoiding hypoglycemia or insulin resistance, reducing and complications

Inactive Publication Date: 2014-06-11
上海坤爱生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there have been no reports on the successful development of stem cell preparations for the treatment of diabetes

Method used

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  • Stem cell preparation for treating diabetes mellitus and preparation method of stem cell preparation
  • Stem cell preparation for treating diabetes mellitus and preparation method of stem cell preparation
  • Stem cell preparation for treating diabetes mellitus and preparation method of stem cell preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: Preparation of human adipose-derived mesenchymal stem cell preparation

[0045] After the qualified fat is mechanically shredded and digested into a single cell suspension by collagenase, the digestion is terminated by adding α-MEM medium or DMEM medium containing fetal bovine serum, and then all the liquid is transferred to a centrifuge tube and centrifuged at 1500rpm 5min, discard the supernatant, add medium to resuspend, adjust the cell density to 1×10 6cells / ml, inoculate the cell suspension at 3ml / per plate in the culture plate of α-MEM medium or DMEM medium containing fetal bovine serum, and when the fibroblasts reach 60-80% fusion, trypsin Digestion, subculture, and re-cultivation to achieve the purpose of purifying and expanding the culture of adipose-derived mesenchymal stem cells. The cells of the 3rd to 8th passages were collected in strict accordance with the cryopreservation steps, frozen and resuscitated for later use.

Embodiment 2

[0046] Example 2: Preparation of Islet-like Cell Preparations Derived from Human Adipose Stem Cells

[0047] After the qualified fat is mechanically shredded and digested into a single cell suspension by collagenase, the digestion is terminated by adding α-MEM medium or DMEM medium containing fetal bovine serum, and then all the liquid is transferred to a centrifuge tube and centrifuged at 1500rpm 5min, discard the supernatant, add medium to resuspend, adjust the cell density to 1×10 6 cells / ml, inoculate the cell suspension at 3ml / per plate in the culture plate of α-MEM medium or DMEM medium containing fetal bovine serum, and digest, Subculture and re-culture to achieve the purpose of purifying and expanding the culture of adipose-derived mesenchymal stem cells. Collect the 3rd to 8th generation cells in strict accordance with the cryopreservation steps, cryopreserve and resuscitate for later use.

[0048] The recovered human adipose stem cells were added to the induction me...

Embodiment 3

[0049] Embodiment 3: Experiment of transplanting adipose-derived stem cell preparations to NOD / SCID rats

[0050] 1) Preparation of adipose-derived stem cell preparation

[0051] a. Prepare adipose-derived human adipose-derived mesenchymal stem cell preparation according to the method of Example 1:

[0052] After the qualified fat is mechanically shredded and digested into a single cell suspension by collagenase, the digestion is terminated by adding α-MEM medium or DMEM medium containing fetal bovine serum, and then all the liquid is transferred to a centrifuge tube and centrifuged at 1500rpm 5min, discard the supernatant, add medium to resuspend, adjust the cell density to 1×10 6 cells / ml, inoculate the cell suspension at 3ml / per plate in the culture plate of α-MEM medium or DMEM medium containing fetal bovine serum, and when the fibroblasts reach 60-80% fusion, trypsin Digestion, subculture, and re-cultivation to achieve the purpose of purifying and expanding the culture...

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Abstract

The invention provides a stem cell preparation for treating diabetes mellitus. The stem cell preparation comprises: stem cells separated and extracted from human blood or tissues and organs and / or islet-like cells induced and differentiated from the stem cells and a cell preservation solution containing 0.5% of heparin calcium, wherein the cell content is 1*10<5> to 5*10<7> / ml. The invention also discloses a preparation method of the stem cell preparation. The method comprises the following steps: (1) separation and purification of the stem cell; (2) in vitro culture of the stem cells or in vitro induction and differentiation of the stem cells to form islet-like cells; (3) diluting the cell obtained in the step (2) by using the cell preservation solution to prepare a stem cell preparation; (4) quality detection of the stem cell preparation. The stem preparation is transplanted into the body and can be differentiated into insulin-secreting cells to secret insulin, and the blood sugar concentration of a person with diabetes is effectively reduced. Thus, a new way is provided for overcoming treatment of the diabetes dependent on the insulin and curing the diabetes once and for all.

Description

technical field [0001] The invention relates 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 mellitus is a type of metabolic disease caused by insufficient insulin secretion or insulin dysfunction, mainly including type I and type II diabetes. In recent years, the incidence of diabetes has been on the rise, and it is getting younger. It is estimated that by 2025, 334 million people worldwide will suffer from diabetes, and diabetes has become a major chronic disease threatening human health. [0003] The current treatment methods for diabetes are mainly traditional treatment methods, including insulin injection, oral hypoglycemic drugs, diet control, and moderate exercise. Clinically, insulin injections are generally used to maintain blood sugar, but insulin injections can neither precisely regulate blood sugar levels in the body nor prevent the occurr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/44A61K35/407A61K35/39A61K35/12A61P3/10C12N5/074C12N5/071A61K35/28
Inventor 刘勇陈彦田齐瀚实
Owner 上海坤爱生物科技股份有限公司
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