Composite mesenchymal stem cell functional collagen scaffold and applications thereof
A collagen scaffold and stem cell technology, applied in medical science, tissue regeneration, prosthesis, etc., can solve problems such as nerve dysfunction that cannot be effectively solved
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Embodiment 1
[0101] Embodiment 1, the preparation of the functional collagen scaffold of composite mesenchymal stem cells
[0102] 1. Isolation and culture of placental mesenchymal stem cells
[0103] 1. Take the placenta of the full-term caesarean section fetus (according to the hospital's regulations and the consent of the family members), and separate the placental chorionic tissue.
[0104] 2. Wash the placental chorionic tissue obtained in step 1 with PBS buffer and cut it into small pieces, digest it with 0.1% (mass fraction) type II collagenase at 37° C. for 90 min, and then use 10% (volume percentage) fetal bovine serum The L-DMEM culture solution was pipetted repeatedly to stop the digestion, and the cell suspension was obtained by passing through a 200-mesh sieve, and the cell suspension was centrifuged at 160 g for 5 min, the supernatant was discarded, and the cell pellet was collected.
[0105] 3. Resuspend the cell pellet obtained in step 2 with L-DMEM medium (containing 10mL...
Embodiment 2
[0127] Example 2. Application of Collagen Scaffold Composite with Mesenchymal Stem Cells to Treat Canine Acute Spinal Cord Transection Injury
[0128] 1. Preparation of canine acute spinal cord transection injury model
[0129] Experimental animals: 1.5-year-old female Beagle dogs (body weight 8-10kg).
[0130] After the experimental animals were anesthetized, they were randomly divided into 6 groups, 6 animals in each group, and the following operations were performed:
[0131] Control group I: The T7-T9 lamina was opened to expose the spinal cord tissue, the spinal cord at the T8 segment was resected by 5 mm, no collagen scaffold was transplanted, and the dura mater was tightly sutured.
[0132] Control group II: Open the T7-T9 lamina to expose the spinal cord tissue, resect the spinal cord at the T8 segment 5 mm, transplant the 5 mm long and 5 mm total diameter collagen scaffold bundle prepared in Step 3 of Example 1, and tightly suture the dura mater.
[0133] Experiment...
Embodiment 3
[0161] Example 3. Application of Collagen Scaffold Composite with Mesenchymal Stem Cells to Treat Dogs with Old Spinal Cord Transection Injury
[0162] 1. Preparation of canine old total transection injury model of spinal cord
[0163] Experimental animals: 1.5-year-old female Beagle dogs (body weight 8-10kg).
[0164] 1. After anesthetizing the experimental animals, open the T7-T9 lamina to expose the spinal cord tissue, resect the spinal cord at the T8 segment by 5 mm, and suture the dura mater tightly.
[0165] 2. At the 9th week (after 2 months) after completing step 1, the experimental animals were anesthetized and randomly divided into 6 groups with 7 animals in each group, and the following operations were performed:
[0166] Control group I: The skin was cut along the original incision, and the epidural and subdural scar tissue was excised under a microscope until the normal spinal cord tissue at both ends was exposed. No collagen scaffold was transplanted, and the du...
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