A cell preparation for treating inflammatory bowel disease
A cell preparation and cell technology, applied in cell culture active agents, animal cells, vertebrate cells, etc., can solve the problems of adverse reactions of patients, affecting the quality of life of patients, and limited efficacy in critical cases
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Embodiment 1
[0089] Example 1 Obtaining Umbilical Cord Wharton Section Mesenchymal Stem Cells
[0090] 1. Fetal umbilical cord acquisition
[0091] The umbilical cords of full-term newborns without congenital diseases were taken; the mothers had no infectious diseases such as hepatitis, syphilis, and AIDS, and the mothers and their families gave informed consent to the use of umbilical cords in the experimental research.
[0092] 2. Obtaining mesenchymal cells from the Wharton section of the umbilical cord
[0093] In a sterile laboratory bench, the umbilical cord was repeatedly washed with phosphate buffered saline (PBS) to remove residual blood. Cut the umbilical cord into small sections of 2-3cm with sterile surgical instruments, cut the umbilical cord longitudinally, remove the umbilical artery, umbilical vein and amniotic membrane, and cut the Wharton area into 0.5-1mm 3 Little pieces left and right. Obtain primary umbilical cord mesenchymal cells by tissue block culture method: sp...
Embodiment 2
[0094] Example 2 Obtaining Umbilical Cord Wharton Section Mesenchymal Stem Cell Preparation
[0095] The above-mentioned low-serum medium was used to culture Example 1 to obtain umbilical cord Wharton interval mesenchymal stem cells. Specifically, the human umbilical cord mesenchymal stem cells isolated by the tissue block method were cultured in the above-mentioned low serum medium, and when the cells were about 80% confluent, they were digested with 0.25% trypsin for 1-2 minutes, and most of the cells were used for expansion. In culture, a small fraction of cells is used to identify markers of stem cells. The expression of CD29, CD44, CD90, CD105, CD31, CD45, CD34 and HLA-DR was detected by flow cytometry (after the cells were incubated with the above corresponding antibodies, the excess antibodies were washed away, the cells were resuspended in PBS, and the The LSRFortessa instrument of BD Company detects the positive rate of each index above), and the results are as follo...
Embodiment 3
[0096] Example 3 Obtaining CD126-positive Umbilical Cord Wharton Interval Mesenchymal Stem Cell Preparation
[0097] The umbilical cord Wharton section mesenchymal stem cell preparation obtained in Example 2 was used to incubate with the CD126 antibody for 20 min at 4° C., to wash away excess antibody, and to resuspend the cells in PBS. Using CD126 as a sorting marker, CD126-positive cells and CD126-negative cells were aseptically sorted by flow cytometry, and the sorting results were as follows: figure 2 As shown, a CD126-positive umbilical cord Wharton mesenchymal stem cell preparation was obtained, and its CD126 ratio was 99%; a CD126-negative umbilical cord Wharton mesenchymal stem cell preparation was obtained, and its CD126 ratio was 0.9%.
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