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Ovary cell microcapsule

A technology of ovarian cells and microcapsules, applied in the biological field

Inactive Publication Date: 2008-07-30
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Whether ovarian cells in vitro can continuously secrete estrogen, whether ovarian cells can continue to secrete estrogen after microencapsulation by microencapsulation technology, and whether they can continuously secrete estrogen after transplanting into the abdominal cavity of mice, and whether it can be effective To prevent the decline of various vital organs of the body (this is a difficult problem that current hormone replacement therapy cannot solve), there has not been any report at home and abroad

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 In vitro culture of ovarian cells

[0045] Under aseptic conditions, the bilateral ovaries of the mouse were removed, the surrounding fat tissue was removed, rinsed with cold PBS, the ovarian tissue was cut into small pieces, and digestive solution (0.2% collagenase I and 0.25% trypsin, mixed in a ratio of 1:1) was added, placed in Digest in a constant temperature water bath shaker at 37°C, add D / F12 culture medium containing 10% FBS after 30min to stop digestion, centrifuge at 179g for 10min, remove the supernatant, wash twice with serum-free culture medium (centrifugation, 179g 10min), gently Pipette to make a cell suspension, filter through a 60-mesh cell sieve, and filter the filtrate at 1×10 6 The density is inoculated at 25cm 2 Add D / F12 (HyClone Company) full culture medium to the culture flask (10% FBS, 1% penicillin-streptomycin are added to the D / F12 culture medium, pH 7.2-7.4), and place at 37°C in 5% CO 2 Incubator cultivation. After the cells w...

Embodiment 2

[0046] Example 2 Preparation of microencapsulated ovarian cells

[0047] Mix the ovary cells of 1-2 passages (without germ cells) with 1.5% sodium alginate to make a cell suspension, and the final cell density is 2×10 6 . Drop the cell suspension into gently agitated 1.1% CaCl with a No. 4 needle 2 , making it form gel beads, adding 0.05% polylysine for 6 minutes. 1% CaCl 2 Wash the gel beads twice, then add 0.15% sodium alginate for 5 minutes. 1% CaCl 2 Wash the gel beads with physiological saline, add 0.05mol / L sodium citrate (pH 7.4, relative molecular weight 249.1), and act for 6 minutes to liquefy the capsule core. After washing with normal saline, wash with culture medium several times, add D / F12 full culture medium, 37°C 5% CO 2 Cultured in an incubator, ready for use. Each microcapsule is about 2mm in diameter and contains about 2×10 4 ovarian cells.

Embodiment 3

[0048] Example 3 Determination of endocrine function of microencapsulated ovarian cells

[0049] The radioimmunoassay method (using Tianjin Jiuding Medical Bioengineering Co., Ltd. progesterone and estrogen radioimmunoassay kit, operated according to the instructions) was used to measure the sex hormone levels secreted by ovarian cells cultured in vitro and microencapsulated ovarian cells.

[0050] Results: Ovarian cells cultured in vitro ( figure 1 ) has endocrine function, microencapsulated ovarian cells ( figure 2 ) compared with ovarian cells cultured in vitro at the same period (Table 1), indicating that the microcapsules have a good permeability effect, and theca cells and granulosa cells in the microcapsules can still continue to maintain synergy, synthesize and secrete Estrogen, progesterone. The ovarian cells packed in the microcapsules were broken and cultured again, and the ovarian cells released from the microcapsules could still adhere to the wall rapidly and g...

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Abstract

The invention provides an ovarian cell microcapsule, which adopts alginate-poly-lysine- alginate (APA) to encapsulate ovarian cells and prepare a microcapsule containing ovarian cells inside. The ovarian cells in the microcapsule consists of ovarian granulosa cells and theca cells, which has endocrine function; undifferentiated ovary inner mesenchymal cells and connective tissue cells. If allogenously plant the microcapsule of the invention, the endocrine cells in the microcapsule can constantly secret sex hormone in body, and the endocrine function can be under the control of neuroendorine of the body to reach the aim of personalized supplementing the inefficiency of sex hormone endocrine. The allogenous plant of ovarian cell microcapsule has positive prevention and treatment effects to body degenerative lesion. Experiment shows that the invention can alleviate the loss of bone quantity and presents positive effect to the prevention and treatment of osteoporosis; meanwhile, the invention can affect the biological activity of islet Alpha cell and Beta cell, and regulate the endocrine of insulin and glucagons.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an ovarian cell microcapsule. Background technique [0002] The ovary is an important reproductive endocrine organ in women, and it has obvious age-related changes. The decline of ovarian function begins at the age of 30, and begins at the menopause. The ovarian function declines rapidly, and the secretion of estrogen drops sharply (the estrogen level after menopause drops by 90% compared with the peak period), which leads to a series of organ dysfunction in the body. Especially the cardiovascular, bone, brain and skin, which are closely related to estrogen, are prone to diseases after menopause. With the extension of human life and the improvement of living standards, the goal of human existence is not only to "live", but also to "live well". Therefore, hormone replacement therapy is used clinically to correct the dysfunction of the body caused by ovarian function decl...

Claims

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Application Information

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IPC IPC(8): A61K35/54A61K8/98A61K9/50A61K47/36A61P15/12A61P43/00A61Q19/00C12N5/08
Inventor 史小林郭晓霞许晴翁静路欣梁元晶
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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