Culture method for enhancing secretion of vascular endothelial growth factors by umbilical cord mesenchymal stem cells
A technology of mesenchymal stem cells and growth factors, applied in the field of biomedicine, can solve the problems of no efficient culture methods for mesenchymal stem cell growth factors, and achieve excellent therapeutic effects, high feasibility, and reasonable process design
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0050] The present embodiment provides a culture method for enhancing the secretion of vascular endothelial growth factor from umbilical cord mesenchymal stem cells, which includes the following contents:
[0051] 1. Extraction of umbilical cord mesenchymal stem cells.
[0052] (1.1) Complete the sample receipt within 24 hours of umbilical cord collection, transfer it to the laboratory and place it in a biological safety cabinet;
[0053] (1.2) Rinse 6-9 times with normal saline containing penicillin-streptomycin 100U / mL, then strip the umbilical vein and umbilical artery and remove the epidermis;
[0054] (1.3) Continue to rinse 3 times with normal saline containing penicillin-streptomycin 100U / mL to avoid blood residue, and then use sterile straight-tip scissors to cut the umbilical cord Wharton's glue to 3mm tissue pieces;
[0055] (1.4) Attach 2ml tissue block to 150cm 2 On the wall of the culture bottle, placed in the opening of the safety cabinet for 2 hours, then slow...
Embodiment 2
[0079] On the basis of Example 1, the excitation time of UC-MSCs was changed.
[0080] The pretreatment method of cells and the detection method of growth factor are the same as the method in Example 1, the difference is that the excitation time of the cells is changed. figure 2 shown.
[0081] Table 2 Different excitation times
[0082]
[0083] as attached figure 2 As shown, in the experiment of changing the time when UC-MSCs were stimulated, after adding the stimulating factor, the amount of VEGF secreted by UC-MSCs was the highest after 48 hours of stimulation.
Embodiment 1
[0084] The content of VEGF in Example 1 was determined by enzyme-linked immunosorbent assay kit at 75cm 2 The UC-MSCs were cultured by adding stimulating factors to the culture flask according to the set conditions, washed with PBS to remove the stimulating factors, and then cultured in the medium without stimulating factors for 12 h. Subsequently, the medium was taken out and centrifuged at 300G for 5 min, and the supernatant was taken to measure the content of VEGF with an ELISA kit.
[0085] According to the excitation conditions determined in Example 1, a new batch of cells was prepared, the expression of VEGF gene was detected by quantitative PCR, the total RNA in cells was collected by RNA extraction kit, and then cDNA synthesis reverse transcription kit was used The RNA was reverse transcribed into a cDNA template, and a quantitative PCR reaction system was prepared using a PCR quantitative reaction kit. Then, the gene expression of UC-MSCs was detected by a real-time f...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More