Stem cell preparation of stem cell composition based on based on ovarian targeting

A stem cell preparation and composition technology, applied in the field of stem cells, can solve the problems of increasing the risk of breast cancer and autoimmune diseases, increasing the risk of heart disease and stroke, etc., and achieve stable in vitro expansion, no toxic side effects, and no ethical and moral issues Effect

Inactive Publication Date: 2021-03-16
广东香雪干细胞再生医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this therapy can relieve the clinical symptoms of POF, HRT cannot fundamentally repair the damaged ovary and restore ovarian function.
In addition, studies have shown that long-term HRT treatment increases the risk of heart disease and stroke, and may increase the risk of breast cancer and autoimmune diseases

Method used

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  • Stem cell preparation of stem cell composition based on based on ovarian targeting

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation and Collection of Fired Dry Cells

[0031] 1. Collect the placenta of a healthy woman voluntarily donated, put it in a large sterile stainless steel equipment basin, pour the appropriate amount of 1 × double anti-physiological saline, clean the placenta, remove the umbilical cord and amniotic membrane, collect the inner layer of the amniotic membrane The connective tissue of the mother surface is placed in a new sterile glass dish, cleaned with physiological saline, removes extra blood and placental lobes, cleans 3-4 times, then use scissors with scissors Cut into small pieces, followed by tissue block: 0.25% trypsin volume = 3: 5, trypsin, 37 degrees, 150-200 rpm, digestion for 30 minutes.

[0032] 2. After digestion, remove the tissue block for 3 times with physiological saline, and thoroughly remove the residual trypsin, then add the tissue block plus 125ml reservoir, add 20 ml of 0.5% I collagenase, add DMEM high sugar. The medium is deposited to 1...

Embodiment 2

[0036] Example 2 Preparation and Collection of Hematophyll Stem Cells

[0037] 1. Collecting the blood-rich young girl volunteer donated blood, mixing the blood and equivalent physiological saline to obtain a blood dilution, separated by a density gradient method, and slowly add blood dilute to a 15 mlfoll lymph isolate In the 50 ml centrifuge tube, the upper layer of the separation solution was added, and 30-40 ml of blood dilute, 800 g centrifuge was centrifuged for 20 min, and the lifting speed was 0.

[0038] 2. After centrifugation, use the bus to absorb the white film layer in the middle of the white film in the new 50 ml of centrifuge tube, resuspended with physiological saline, 500g centrifugation for 5 min, then use blood-free mesenchymal stem cells complete medium heavy, inoculated into the culture In the bottle, inoculation, according to 4000-6000cm 2 , Placed in 37 degrees, 5% CO 2 Culture in the incubator.

[0039] 3. After 48 hours, the liquid is started, and the cel...

Embodiment 3

[0041] Example 3 Preparation of Stem Cell Preparation

[0042] Examples 1 and Example 2 were collected into placental mesenchymal stem cells and blood-repellent stem cells, sterile detection and endotoxin were negative, and cellular viability reached 90%, and cell fluid phenotypes must conform to dry cytokines. Type standard (CD73, CD90, CD105 ≥ 95.0%; CD34, CD45, HLA-DR ≤ 2.0%).

[0043]1. A number of 1 × 10 8 Placental mesenchymal stem cell suspension and quantity is 1 × 10 8 The blood mixed stem cell suspension is fully mixed, and 500 g is centrifuged for 5 min.

[0044] 2. Abandon the clear cells, and then rehacked with physiological saline, and 300 g centrifuge for 5 min.

[0045] 3. Abandon the sewen, resuspending the cells with 1.7 ml of physiological saline and 0.7 ml 20% albumin, and then filtered with 40 μm sterile cell filter, and 2 ml of stem cell preparation was made.

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Abstract

The invention relates to the technical field of stem cells, in particular to a stem cell preparation of a stem cell composition based on ovarian targeting. The invention discloses the stem cell composition which consists of placenta mesenchymal stem cells and menstrual blood mesenchymal stem cells. According to the stem cell composition, the placenta mesenchymal stem cells and the menstrual bloodmesenchymal stem cells are combined for use, so that the estrogen level can be effectively reduced, ovarian injury can be repaired, the ovarian function can be recovered, and the treatment effect on premature ovarian failure is remarkably superior to that of single use of the placenta mesenchymal stem cells or the menstrual blood mesenchymal stem cells. The combination of the placenta mesenchymalstem cells or the menstrual blood mesenchymal stem cells has a synergistic effect. The two mesenchymal stem cells in the stem cell composition are wide in source, stable in in-vitro amplification, free of ethical morality problems, low in immunogenicity, safe to use and free of toxic and side effects.

Description

Technical field [0001] The present invention relates to the field of stem cells, and more particularly to a stem cell formulation based on ovarian targeted stem cell composition. Background technique [0002] Prematureovarianfailure, POF is a phenomenon before the 40-year-old closing of the ovarian failure. It is a disease in a closing, infertility, estrogen lack, follicular decrease and increased gonadotropin, and has a series of low estrogen symptoms such as: hot tide sweat, facial flushing, low-quality, and severely affect women Physical and mental health. In addition, POF has increased risk of osteoporosis, cardiovascular disease and dementia. POF accounts for 1-3% in women, and the incidence in recent years has a rise. [0003] The cause of POF is complex, has not fully clarified, but related to autoimmune response, infection, genetic factors, chemotherapy, radiotherapy, surgery and other treatment effects and endocrine dysfunction. At present, the most common treatment of P...

Claims

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Application Information

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IPC IPC(8): A61K35/50A61K9/08A61K47/42A61P15/08A61P15/00A61K35/28
CPCA61K35/50A61K35/28A61P15/08A61P15/00A61K9/0034A61K9/08A61K47/42A61K2300/00
Inventor 张玲洁陈育广张林艳
Owner 广东香雪干细胞再生医学科技有限公司
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