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Stem cell preparation as well as preparation method and application thereof

A technology of stem cell preparation and qualitative stem cells, which is applied in the direction of pharmaceutical formulas, medical preparations containing active ingredients, peptide/protein components, etc., can solve the problems of small number of autologous bone marrow, unfavorable genome stability, etc., and achieve strong in vitro differentiation ability, Short preparation time and enhanced VEGF expression

Inactive Publication Date: 2015-12-23
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the stem cell therapy of femoral head necrosis, the commonly used seed cells are bone marrow mesenchymal stem cells. However, due to the extremely small amount of autologous bone marrow, it is difficult to culture and expand cells to a certain therapeutic amount in vitro, and it is not conducive to maintaining their genome stability.

Method used

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  • Stem cell preparation as well as preparation method and application thereof
  • Stem cell preparation as well as preparation method and application thereof
  • Stem cell preparation as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Example 1. Acquisition of adipose-derived mesenchymal stem cells

[0035]Take 50mL of fat, wash it twice with PBS, add 0.3%-0.5% collagenase I to digest it for 50min, wash it once with PBS, add complete medium to resuspend the cell pellet, inoculate it into a culture dish and put it in a carbon dioxide incubator for cultivation After 48 hours, the medium was changed; when the cell confluence reached 80%-90%, subculture was carried out; the third to fifth generation adipose stem cells were washed twice with PBS, digested with trypsin for 2 minutes, and then digested with The complete medium was digested, and repeatedly blown and mixed with a pipette gun until the cells fell off; the stem cell suspension was transferred to a 50mL centrifuge tube, and centrifuged at 1500rpm for 5min; at the same time, the cells were counted to ensure that the total number of cells was 1.0×10 8 -5.0×10 8 1; after washing twice with normal saline, resuspend the cells with albumin in normal ...

Embodiment 2

[0036] Example 2, Quality Evaluation of Adipose-derived Mesenchymal Stem Cells

[0037] The cell morphology, cell viability and surface antigen of the adipose-derived mesenchymal stem cells prepared in Example 1 were detected respectively, and the cells without any antibody were used as the control cell group, and the results are shown in Figure 1-3 .

[0038] Depend on figure 1 The results can be seen, observed under an inverted microscope, the stem cell preparation prepared in Example 1 of the present invention has strong cell proliferation ability, fast growth rate, uniform shape and size, strong refraction of cells, and grows in a radial to spindle-shaped orderly arrangement, which conforms to Adipose-derived mesenchymal stem cell properties.

[0039] Depend on figure 2 The results showed that after staining with trypan blue, most of the adipose-derived stem cells were still translucent without being stained blue, and their viability was 96%.

[0040] The recognized ...

Embodiment 3

[0042] Embodiment 3, PRP preparation

[0043] Centrifuge 10mL of autologous whole blood at 200g-400g for 5min-10min, remove the upper plasma and discard the red blood cells; centrifuge the upper plasma at 1200g-1600g for 5min-10min, keep 1mL of plasma, discard the rest, resuspend the pellet, and obtain PRP.

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Abstract

The invention relates to the field of stem cells, and discloses a stem cell preparation as well as a preparation method and an application thereof. The stem cell preparation comprises adipose derived stem cells and is wide in source, high in in-vitro differentiation capacity, high in safety and good in treatment effect. The stem cell preparation also comprises high-concentration platelet-rich plasma, so that the activity of the adipose derived stem cells in the preparation can be improved, and the treatment of the adipose derived stem cells on femoral head necrosis can be promoted. The preparation method is short in preparation time and applicable to the mass production. The stem cell preparation can promote the activity of the bone cells and is favorable for the generation of new bones and capable of enhancing the VEGF expression and enhancing the generation of local necrotic blood vessels, a complete blood vessel net is formed, and the stem cell preparation can be used for preparing a drug for treating the femoral head necrosis.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a stem cell preparation and its preparation method and application. In particular, it relates to a stem cell preparation containing adipose stem cells, a preparation method thereof, and an application in preparation of medicine for treating femoral head necrosis. Background technique [0002] Femoral head necrosis, also known as aseptic necrosis of the femoral head, is a challenging and destructive disease in bone and joint diseases, and it is also a common and intractable disease in the field of orthopedics. Femoral head necrosis can occur at any age, but it is most common in the age group of 31-60. There is no gender difference. At first, it usually manifests as dull pain or dull pain in the hip joint or its surrounding joints, which aggravates after activities, and further development can lead to dysfunction of the hip joint. Necrosis of the femoral head seriously affects the patien...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K35/16A61K35/19A61K38/38A61P19/08
Inventor 王一飞陈海佳葛啸虎马岩岩王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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