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Neural stem cell phospholipid membrane microcapsule and preparation method thereof

A technology of neural stem cells and phospholipid membranes, applied in the field of neural stem cell phospholipid membrane microcapsules and its preparation, phospholipid membrane microencapsulated neural stem cells, can solve the problems of limited application, large differences in tissue components, foreign body reactions, etc., and achieve the preparation process Simple, less toxic and side effects, good safety effect

Active Publication Date: 2021-12-03
THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the field of brain tissue repair and functional replacement, the application of pure alginate-polylysine microcapsules is extremely limited, and it is not suitable for nerve repair in diseases such as cerebral infarction: on the one hand, because alginate, polylysine Lysine is extracted from brown algae and actinomycetes respectively, and its biocompatibility is not good due to the large difference with human tissue components, which may cause inflammation, tissue fibrosis, etc., and then affect neuron function
Based on the inflammatory response induced by pathological conditions (such as cerebral infarction), transplantation of simple alginate-polylysine microcapsules is more likely to cause foreign body reactions; on the other hand, simple alginate microcapsules are stable It is used to protect the cells in the capsule (such as islet cells, etc.) from immune rejection for a long time, while the long-term encapsulation of neural stem cells in microcapsules may block the surface receptors of stem cells, so that their chemotactic activities in vivo and Signaling connections are disturbed so that they do not integrate well with brain tissue

Method used

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  • Neural stem cell phospholipid membrane microcapsule and preparation method thereof
  • Neural stem cell phospholipid membrane microcapsule and preparation method thereof
  • Neural stem cell phospholipid membrane microcapsule and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Preparation of neural stem cells microencapsulated by phospholipid membrane

[0048] 5% (w / v) of L-α-phosphatidylcholine was added to 2% (w / v) alginate solution, then the cultured neural stem cells were added to obtain a neural stem cell final concentration of 10. 6 / ml cell suspension; load cell suspension into a microcapsule electrostatic droplet generator, a flow rate of 10 ml / h, a pressure of 100 kPa, a calcium chloride solution in 100 mmol / L, and a reaction for 10 minutes. It was rinsed three times with 0.9% physiological saline, then added to a polyliotine solution of 0.1%, the reaction time was 10 minutes, and after rinse with 0.9% saline, it was added to 55 mmol / l sodium citrate solution. A neural stem cell suspension forming a phospholipid-alginate-multi-lysine microencapsulated.

[0049] With phosphatidylcholine as a raw material, chloroform is a solvent, formulated concentration of 2 mg / ml phospholipid solution; apply a phospholipid solutio...

Embodiment 2

[0051] Example 2 Survival of microencapsulated neural stem cells in the OGD model

[0052] To construct an OGD model as follows: Discharge Sugar DMEM / F12 Culture, adjust the number of cells of each experiment to 1 × 10 5 / ml concentration, inoculated to 96-well plate, perfoated by 100 μL; place cells were placed in an oxygen-free breeding box (85% nitrogen, 10% carbon dioxide, 5% hydrogen), 37 ° C for 2 hours; after 2 hours, half Exchange method and use the original cell culture solution instead of a sugar-sugar DMEM / F12 culture solution, and the cells return to the regular oxygen incubator. After returning to normal cultivation of 0, 24, 48 hours, the CCK8 statutory detection and contrast the number of cells of the experimental group of each phase of the phase, as follows: collecting the cells, PBS gently rinsing 3 times, transfer to new 96 holes The plate was added to 100 μl of medium, and then 10 μl of CCK8 reagent was added, 1 hour at 37 ° C, the enzyme laminator 450 nm w...

Embodiment 3

[0053] Example 3 TTC dyeing, detecting the effects of neuronal cells on the microencapsimization of phospholipid membranes on cerebral infarction

[0054] The MCAO model is constructed as follows, and adult C57BL / 6 mice, 1% pentobarbital sodium solution (0.01 mg / g body weight) is conventional anesthesia. Cut the neck skin, expose the right neck at the carotid artery, the intraocular artery, the neck artery, and the carotid artery is separated. The long-necked artery is ligated, and the proximal end is single. The neck internal artery row, respectively, the intimate end of the cervic artery and the neck of the heart, temporarily block the blood flow, then cut all the mouth in the neck artery single knot, insertion from the incision Tie. External artery, release the intra-neck arterial single routine. Adjusting the insertion direction, pointing to the intraocular direction, circulating the blood vessel insertion into the middle of the cerebral artery, the incision is covered wit...

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Abstract

The invention discloses a neural stem cell phospholipid membrane microcapsule and a preparation method thereof. The neural stem cell phospholipid membrane microcapsule is mainly prepared from neural stem cells, sodium alginate, phosphatidylcholine and polylysine. According to the phospholipid membrane microcapsule; phospholipid widely existing in brain tissue is introduced into a traditional alginate-polylysine microcapsule, so that a tissue engineering microcapsule which is extremely good in biocompatibility and integration with the brain tissue is constructed; and animal experiments prove that the neural stem cell phospholipid membrane microcapsule can be applied to nerve repair after brain tissue injury, and the disability rate is reduced.

Description

Technical field [0001] The present invention belongs to the field of biotechnology medicine, in particular to a neural stem cell phospholipid membranes and preparation method microcapsule or microencapsulated one phospholipid membrane of neural stem cells. Background technique [0002] Physiological basis of brain tissue damage after high morbidity of pathological brain tissue neuronal necrosis, and therefore nerve repair after brain injury is the key. Unfortunately, currently no effective way to promote nerve repair major clinical treatment is symptomatic of physical rehabilitation for neurological deficit symptoms. In the leading basic research, reported in the literature despite promote repair by means of regulation of gene expression, but also far away from clinical use. In contrast, stem cell transplantation therapy is even feasible for the brain nerve repair after tissue injury has brought a ray of hope. [0003] The body's own neural stem cells can be activated in patholog...

Claims

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Application Information

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IPC IPC(8): A61K9/50A61K47/36A61K47/34A61K35/30A61K47/24A61P25/00A61P9/10A61P7/04
CPCA61K9/5036A61K9/5031A61K35/30A61K47/24A61P25/00A61P9/10A61P7/04
Inventor 徐瑞段春梅何胜荣杨清武陈晓燕刘舒
Owner THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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