Exosome derived from human pluripotent stem cells, preparation based on exosome and use of exosome and preparation

A technology of human pluripotent stem cells and exosomes, applied in non-embryonic pluripotent stem cells, artificially induced pluripotent cells, embryonic cells, etc., can solve the problems of bone degenerative diseases and incapacitating diseases, and achieve large-scale applications Value, promoting tissue cell regeneration, and the effect of treating bone diseases

Inactive Publication Date: 2019-04-23
上海睿泰生物科技股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the tumorigenicity of ESCs or iPSCs, they cannot be directly transplanted for disease prevention and treatment
However, th

Method used

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  • Exosome derived from human pluripotent stem cells, preparation based on exosome and use of exosome and preparation
  • Exosome derived from human pluripotent stem cells, preparation based on exosome and use of exosome and preparation
  • Exosome derived from human pluripotent stem cells, preparation based on exosome and use of exosome and preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Culture of human embryonic stem cells (ESCs) and extraction and identification of exosomes

[0090] A layer of embryonic stem cell Matrigel (ESC-Qualified BD Matrigel, BD Sparks, MD, USA), ESCs were moved into the dish, and mTeSR1 serum-free medium (StemCell Vancouver, BC, Canada), in an incubator (37°C, 5% CO 2 , saturated humidity) culture, and collect the culture medium changed every day. Filter the medium through a 0.22 micron pore size filter membrane and centrifuge at 10,000g at 4°C for 30 minutes to remove cell debris; use a 100KD molecular weight ultrafiltration tube and centrifuge (3500g, 15min) to intercept exosomes in the concentrated supernatant to obtain exosomes concentrate; transfer the concentrate to a 30% sucrose / heavy water density pad (1.210 g / cm 3 ), centrifuge at 100,000g at 4°C for 210 minutes, collect the 5ml sucrose / heavy water density pad at the bottom, add PBS to dilute, transfer to an ultrafiltration centrifuge tube with a molecular weigh...

Embodiment 2

[0100] Prevention and treatment effect of exosomes derived from human pluripotent stem cells on osteoporosis

[0101] Prevention experiment: select the 6-month-old SAMP6 rapid aging mouse model, and divide the experiment into two groups, with 10 mice in each group. The experimental group is administered 2-5×10 8 Individual ES-exo or iPS-exo were intervened, and the control group was given the same volume of normal saline. The mice were killed 3 months after the intervention, and the samples were taken for testing.

[0102] Therapeutic experiment: select the 8-month-old SAMP6 rapid aging mouse model, divide the experiment into two groups, 10 mice in each group, and give human 2~9×10 8 Each ES-exo was intervened, and the control group was given the same volume of normal saline. The mice were killed 3 months after the intervention, and the samples were taken for testing.

[0103] Blood was collected from the mice before sacrifice, and the levels of serum tartrate-resistant aci...

Embodiment 3

[0108] Preventive effect of exosomes derived from human embryonic stem cells on avascular necrosis of the femoral head

[0109] The culture of human embryonic stem cells and the extraction of exosomes are the same as in Example 1. In the experiment, 300-320 g male SD rats were used to make a model of hormone-induced necrosis of the femoral head: SD rats were intramuscularly injected with methylprednisolone (40 mg / kg), 3 times a week for 3 weeks. Experimental grouping: model group (hormone-induced necrosis of the femoral head, n=10); ES-exo intervention group (hormone-induced and simultaneous intravenous administration of ES-exo for intervention, n=10); normal control group (given the same dose of PBS, n=10). After all injections were completed, the rats were housed in cages and raised normally for 3 weeks. Then samples were taken to evaluate the neovascularization and osteonecrosis of the femoral head.

[0110] Detect changes in the bone structure of the femoral head by mic...

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Abstract

The invention relates to an exosome derived from human pluripotent stem cells, a preparation based on the exosome and use of the exosome and preparation, and belongs to the technical fields of cell biology, molecular biology and drug research and development. The human pluripotent stem cell exosome or the preparation based on the human pluripotent stem cell exosome has the uses of A, being an additive cultured in vitro for maintaining the stemness of adult stem cells and inhibiting the senility of the adult stem cells; B, being an additive cultured in vitro for maintaining the proliferative activity of various tissue primary culture cells; C, being applied in preparing medicine for treating bone and cartilage degenerative changes; D, being applied in preparing medicine for preventing the bone and cartilage degenerative changes; E, being applied in preparing medicine for treating orthopedic diseases such as osteonecrosis, bone nonunion, fractures, cartilage injuries, tendon injuries, frozen shoulders or tendon adhesion. Compared with the prior art, the exosome secreted by the pluripotent stem cells has powerful functions of promoting tissue cell regeneration, preventing senility andtreating bone diseases.

Description

technical field [0001] The invention belongs to the technical fields of cell biology, molecular biology and drug research and development, and in particular relates to exosomes derived from human pluripotent stem cells, preparations based on the exosomes and uses thereof. Background technique [0002] With the increase of age, the functions of various organs of the human body will gradually age, and cause the occurrence of related diseases. With the aging society in our country, the incidence of cardiovascular and cerebrovascular diseases, bone and joint diseases, nervous system diseases and metabolic diseases have increased significantly, and the harm of senile diseases has become increasingly prominent. At present, there are more than 200 million osteoporotic patients in the world, and 1 / 3 of women over 50 years old have osteoporotic fractures, which have a high disability and mortality rate, seriously threaten public health, and bring great socioeconomic burden. In addi...

Claims

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Application Information

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IPC IPC(8): C12N5/0735C12N5/074A61P19/04A61P19/08
CPCA61K35/545C12N5/0606C12N5/0696C12N2500/84
Inventor 汪泱吴复跃何振东
Owner 上海睿泰生物科技股份有限公司
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