Application of pluripotent stem cell-derived mesenchymal stem cells in treatment of psoriasis
A technology of pluripotent stem cells and mesenchymal stem cells, applied in the application field of mesenchymal stem cells in the treatment of psoriasis, can solve the problems of donor injury, MSCs difficult to standardize, and hinder the clinical treatment of MSCs
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Embodiment 1
[0039] Psoriasis repair with MSCs derived from hESCs.
[0040] 1. Obtain MSCs derived from hESC:
[0041] The hESC cell line H9 was differentiated into MSCs (differentiation medium: K-DMEM + 10% KSR + 0.5% PSG + 0.5% NAA + 8ng / ml bFGF + 4ng / ml PDGF + 1% ITS), by flow cytometry The surface markers CD105, CD90, and CD29 specifically expressed by MSCs were identified to determine the differentiation efficiency of hESCs to MSCs, and their biological characteristics were identified through adipogenic and osteogenic differentiation.
[0042] Morphological observation of H9 embryonic stem cells see figure 1 , hESCs cultured in the feeder-free system grow like clones, with good light transmission, cells in the clones are tightly arranged, and the ratio of nucleoplasm to cytoplasm is high. After being induced to differentiate into MSCs, the cell morphology was as follows figure 2 As shown, it becomes a long fusiform. Through flow cytometric analysis, it was found that hESC-MSCs hi...
Embodiment 2
[0053] Inject hESC-derived MSCs.
[0054] Injection of hESC-derived MSCs may also have similar effects, but the route of treatment is harmful.
[0055] The mice used to establish the mouse psoriasis model were purchased from (Xiamen University Experimental Animal Center). The back hair of the mice was shaved off, and 5% imiquimod (Imiqumod, IMO) ointment was applied daily to create a mouse psoriasis model. All the mice were housed in IVC independent cages, with normal diet and water.
[0056] Treatment began on the day of modeling, and hESC-derived MSCs were injected subcutaneously into the damaged skin every day, and the wound healing was observed on the 7th day.
[0057] Such as Figure 7 As shown, the 7-day corrected PASI score of the MSC-injected treatment group was 5.00±0.47, which was significantly better than that of the control group.
Embodiment 3
[0059] Psoriasis repair with hiPSC-derived MSCs.
[0060] 1. Obtain hiPSC-derived MSCs:
[0061] hiPSCs are cells obtained from healthy people and prepared according to the method reported in the article by Shinya Yamanaka et al. (Cell, 2007, 131, 861-872).
[0062] The hiPSC cells were differentiated into MSCs (differentiation medium: K-DMEM+10% KSR+0.5% PSG+0.5% NAA+8ng / ml bFGF+4ng / ml PDGF+1%ITS).
[0063] The result is as image 3 As shown, the differentiated MSCs have the typical morphological characteristics of MSCs, which are long spindle-shaped and closely arranged cells.
[0064] 2. Establish a mouse skin psoriasis model:
[0065] The mice used to establish the mouse psoriasis model were purchased from (Xiamen University Experimental Animal Center). The back hair of the mice was removed, and imiquimod (Imiqumod, IMO) was applied daily to create a psoriasis model, (the mice were all kept in IVC independent mouse cages, with normal diet and water supply)
[0066] 3....
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