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3D printed artificial ovarium biological stent capable of activating original ovarian follicle and artificial ovarium and application thereof

A primordial follicle and biological scaffold technology, applied in the field of 3D printing biological scaffold preparation, can solve the problems of irreplaceable effects, hormone therapy is not suitable for everyone, difficult to give doses, etc., to achieve rich nutrient supply, shorten activation time, reduce The effect of waiting time

Active Publication Date: 2019-05-14
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although it has a certain effect on improving menopausal symptoms, there are also some disadvantages, such as: hormone therapy is not suitable for everyone, and patients with breast, uterine diseases, diabetes, high blood pressure and other diseases should use it with caution; The amount required by each person is also different, and it is difficult for doctors to give the most suitable dose, and excessive hormones in the body can cause uncontrollable side effects such as endometrial cancer, ovarian cancer, or breast cancer; externally administered hormones are artificial Synthesis, no matter in terms of composition or proportion, it can never replace the effect of self-secreted hormones

Method used

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  • 3D printed artificial ovarium biological stent capable of activating original ovarian follicle and artificial ovarium and application thereof
  • 3D printed artificial ovarium biological stent capable of activating original ovarian follicle and artificial ovarium and application thereof
  • 3D printed artificial ovarium biological stent capable of activating original ovarian follicle and artificial ovarium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Design and preparation of 3D printed bioscaffold

[0056] (1) Combined with computer aided design (CAD), three-dimensional printing technology is used to construct biological scaffolds. The shape of 3D biological scaffolds is selected as circular, rectangular, square, elliptical or irregular, and the internal structure is connected. Pore; pore size (R): 400μm; line stacking angle: 30-120 degrees;

[0057] (2) Prepare 3D printing bio-scaffold printing materials, which include a mixture of biocompatible materials, primordial follicle activators, and primordial follicle development promoters; the biocompatible materials are 3%-5% sodium alginate and 8%- 10% gelatin, the mass ratio of sodium alginate and gelatin is 1:1, it becomes gelatinous at room temperature after mixing. Add 740Y-P 10ug / ml, PA 8uM, PRO 2uM, FSH 100mIU, LH 10mIU, EGF 5ng / ml, ITS (used after 100-fold dilution) to the biocompatible material.

[0058] (3) Print the ovarian stent by 3D printing; ...

Embodiment 2

[0060] Example 2 Design and preparation of 3D printed bioscaffold

[0061] The scaffold prepared in Example 2 is the same as that in Example 1, except for the formulation of the printing material in step (2), which is formulated according to the proportions listed in the table below.

[0062]

[0063]

[0064] Example 2 Obtaining of primordial follicles

[0065] The 3-day-old female mice were killed by cervical dislocation, and after the skin was disinfected, the ovaries were aseptically removed from the dorsal position, and put into the separation medium (L-15+10%FBS+100IU / ml penicillin+100ug / ml chain mycin). The accompanying tissues around the ovaries were removed under a solid microscope, and washed 3 times in the separation solution. Then the ovary was needled with insulin metered injection under the microscope, and the mouse ovary was divided into tissue fragments containing primordial follicles.

Embodiment 3

[0066] Example 3 Activation of primordial follicles

[0067] The activator is an agonist or inhibitor of a signaling pathway related to primordial follicle activation, which belongs to the prior art and can be purchased from the market.

[0068] The cryopreserved or fresh ovarian cortex containing a large number of primordial follicles was minced, incubated and activated in basal culture medium containing different combinations of follicle activators for 0.5 to 6 hours, washed with normal saline three times to remove foreign substances, and then moved into the printing A good bioscaffold was transplanted under the kidney capsule of an adult female rat with bilateral ovariectomy.

[0069] Wherein the basal culture solution formula of the follicle activator is:

[0070]

[0071]

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Abstract

The invention provide a 3D printed artificial ovarium biological stent capable of activating an original ovarian follicle. According to the 3D printed artificial ovarium biological stent, a 3D printing technology is utilized to mix an original ovarian follicle activating agent and hormones and growth factors which can promote the developing of the ovarian follicle into a stent material to be molded through printing, an ovarian cortex fragment which contains the original ovarian follicle is transplanted, after the cortex fragment is transplanted back to the body, the cortex fragment can achievebetter stent protection and more diversified nutrient supplying, the biological stent is gradually degraded in the body to sustainably release the ovarian follicle activating agent, and therefore theoriginal ovarian follicle is sustainably activated. The stent has the advantages that the in vitro activating time of the original ovarian follicle is greatly reduced, a novel researching tool is provided for an revealing an original ovarian follicle activating mechanism, the waiting time of the patient is reduced on the basis that secondary surgery is avoided, and a novel choice is provided forthe clinical treatment of premature ovarian failure and an assisted reproduction technology; after the ovarian follicle is activated, a large number of hormones are secreted to regulate the secretionin the body, and a novel researching direction is provided for improving comprehensive symptoms of menopause and ageing slowing.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the preparation of a 3D printed biological scaffold capable of activating primordial follicles in the ovarian cortex. Background technique [0002] With the rapid development of my country's social economy, the status of women is getting higher and higher, and at the same time, they are also facing increasing work and psychological pressure, and the age of childbearing for women of childbearing age is continuously postponed; Harmful substances are getting closer and closer to life, and the combination of social and environmental factors has caused a large number of reproductive health problems; and since the opening of the second-child policy, there have been more and more elderly women. Therefore, how to make women of super-fertile age who have gradually declined ovarian function successfully complete the pregnancy process has become a major research focus in recent years. [0003] ...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12Q1/02A61P15/08A61P35/00
CPCA61L27/40A61L27/54A61P15/08A61P35/00C12Q1/02
Inventor 张键杨雅莉赵华山
Owner SHENZHEN INST OF ADVANCED TECH
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