Combined cellular preparation and application thereof

A cell preparation and preparation technology, applied in the field of stem cells, can solve the problems of slow expansion, few stem cells, aggravating the disease, etc., and achieve the effects of fast expansion, restoration of lower limb blood flow, and simple operation.

Inactive Publication Date: 2017-10-17
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage is that the amount of bone marrow-derived stem cells is small and the expansion speed is slow, which limits its application.
In addition, harvesting bone marrow places a huge physical burden on the patient, which may exacerbate the condition

Method used

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  • Combined cellular preparation and application thereof
  • Combined cellular preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Preparation of Umbilical Cord Mesenchymal Stem Cell Preparation

[0021] (1) Under sterile conditions, take out the fresh umbilical cord that has been away from the mother for no more than 2 hours, and screen whether the parturient is carrying infectious disease pathogens (negative for hepatitis B surface antigen, hepatitis C antibody, syphilis antibody, and AIDS antibody), and place the fresh umbilical cord in place. Transport back to the laboratory in PBS containing double antibodies in a 4°C incubator;

[0022] (2) Clean the fresh umbilical cord with aseptic PBS, sterilize with 75% ethanol, and then clean it with PBS;

[0023] (3) Cut the umbilical cord into 20mm 3 and inoculated evenly in the culture dish at intervals of 0.5-1 cm, slowly added DMEM-F12 complete medium (purchased from GIBCO), and then placed at 37°C, CO 2 Static culture in an incubator with a concentration of 5%; the amount of complete medium added is 50-100 μl / cm 2 ;

[0024] (4) After...

Embodiment 2

[0027] Example 2 Preparation of NKT cell preparation

[0028] (1) Collect 40mL of peripheral blood, mix it with normal saline at a volume ratio of 1:1, then slowly add the mixed solution into the lymphocyte separation medium (Axis-Shield company), wherein the mixed solution and the lymphocyte separation medium The volume ratio is 2:1; then, centrifuge at 700g for 20min;

[0029] (2) The centrifuge tube after centrifugation is mononuclear cell layer (PBMC), lymphocyte separation liquid layer and red blood cell layer from top to bottom; collect the mononuclear cell layer, resuspend and wash with normal saline, and centrifuge at 500g for 10min, so Repeat washing 2 times;

[0030] (3) Resuspend the mononuclear cells with RPM1640 basal medium, add 1×10 6 Cells / ml density was inoculated in culture flasks, cell growth factors IL-2 and IL-15 were added, and then placed at 37°C with a concentration of 5% CO 2 Cultivate in an incubator; wherein, the final concentration of IL-2 is 500...

Embodiment 3

[0039] According to the classic Wagner grading method, the degree of injury of diabetic foot patients can be divided into 5 grades, including grade 0: feet at risk of developing foot ulcers, no open skin lesions; grade 1: ulcers on the surface, no clinical infection; 2 Grade: Deep ulcer infection lesions, often combined with soft tissue inflammation, without abscess or bone infection; Grade 3: Deep infection, accompanied by bone tissue lesions or abscess; Grade 4: Bone defect, partial gangrene of toes and feet; Grade 5 : Gangrene of most or all of the foot.

[0040] Include 30 diabetic foot patients of grade 2 and grade 3 respectively, and randomly divide them into 3 groups, with 10 diabetic foot patients of grade 2 and grade 3 in each group, followed by the combination preparation group, control group 1 and control group 2, according to the following method For treatment:

[0041] Combined preparation group: the umbilical cord mesenchymal stem cell preparation of Example 1 a...

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Abstract

The invention discloses a combined cellular preparation and an application thereof, and belongs to the technical field of the stem cells. The combined cellular preparation comprises an umbilical cord mesenchymal stem cell preparation and a NKT cellular preparation. An umbilical cord is used as the tissue source of the mesenchymal stem cells, the operation is simple, and the stem cells are easily obtained. A mononuclear cell in peripheral blood is used for inducing to form the NKT cells, and the preparation has the obvious function of improving the organism immunity, and eliminating the body infection. The wound healing of a diabetic foot patient is promoted, and the combined preparation of two preparations has the synergistic interaction function, and is capable of further promoting the recovery of the patient.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to a combined cell preparation and its application. Background technique [0002] Diabetic foot refers to due to comprehensive factors such as diabetic vascular disease, neuropathy and infection, neuropathy leads to lower limb protection function, and macrovascular and microvascular disease causes insufficient arterial perfusion to cause microcirculation disturbance, resulting in foot pain and deep skin ulcers. Even the diseased state of gangrene of the limbs. Diabetic foot is a serious complication of diabetes. It is one of the important causes of disability and even death of diabetic patients. It not only causes pain to patients, but also adds a huge economic burden to them. [0003] The main treatment for diabetic foot is stem cell transplantation. Stem cell transplantation to treat diabetic foot is mainly based on the principle that stem cells can differentiate ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P3/10A61P9/14A61K35/17
CPCA61K35/28A61K9/0019A61K35/17A61K2300/00
Inventor 陈海佳葛啸虎王一飞李丽娟王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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