Cell therapy composition for treating osteoarthritis and cartilage defects
A technology for cell therapy and osteoarthritis, applied in the field of stem cell cell therapy compositions, can solve problems such as inability to repair cartilage tissue, achieve high survival rate, and solve the effect of low survival rate in joint cavity
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Embodiment 1
[0026] Example 1 Extraction and cultivation of adipose tissue-derived stem cells
[0027] This example is used to illustrate the method for extracting and culturing stem cells derived from adipose tissue, and the specific steps are as follows.
[0028] Abdominal adipose tissue from patients or fat donors was taken, washed 3 times with PBS, digested with the same volume of 0.15% type I collagenase for 1 hour, diluted with PBS and centrifuged at 1800r / min for 10 minutes to obtain a precipitate; the precipitate was mixed with PBS Suspend, add erythrocyte lysate, remove erythrocytes, centrifuge at 1200r / min for 5min to obtain a precipitate; wash the precipitate with PBS again, suspend and inoculate it into culture medium with 10% FBS L-DMEM medium (or human mesenchymal stem cell serum-free medium) dish and at 37°C, 5% CO 2 After 24 hours, most of the stem cells have adhered to the wall, and the unattached cells are discarded, and new medium is added to continue the culture; the m...
Embodiment 2
[0030] Example 2 Isolation and cultivation of bone marrow-derived stem cells Stem cells
[0031] This example is used to illustrate the method for extracting and culturing bone marrow-derived stem cells, and the specific steps are as follows.
[0032]Collect the bone marrow from the patient or a bone marrow donor, centrifuge at 1200r / min for 10min, add red blood cell lysate to the pellet, lyse the red blood cells, and centrifuge at 1200r / min for 5min. The cells were suspended and inoculated into culture dishes with 10% FBS in L-DMEM medium (or human mesenchymal stem cell serum-free medium) and incubated at 37°C, 5% CO 2 and cultured under standard conditions of saturated vapor pressure. After 24 hours, most of the stem cells had adhered to the wall, the unattached cells were discarded, and new medium was added to continue culturing. The culture medium was changed once a day for the first two days of culture, and the culture medium was changed every other day in the later per...
Embodiment 3
[0035] This example is used to illustrate the composition and preparation method of a cell therapy composition for treating osteoarthritis and cartilage defect.
[0036] Party:
[0037] Xanthan gum saline solution 2mL
[0038] (average molecular weight 1.4 million, concentration 10000μg / ml)
[0039] Stem cells derived from adipose tissue (primary to 10th passage) 2×10 7 cells
[0040] Preparation method: take the monolayer cultured stem cells from primary to 10th passage in Example 1, wash them with PBS for 3 times, then digest them with 0.25% trypsin-0.02% EDTA digestion solution for 3 minutes, add an appropriate amount of L-DMEM containing 10% FBS Culture medium (or serum-free medium for human mesenchymal stem cells), gently blow the cells to make them fall off, suspend into a single cell suspension, count, centrifuge at 1200r / min for 3min, discard the supernatant, and wash twice with PBS, Press at least 1×10 7 Add appropriate amount of sterilized xanthan gum solution o...
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