Tissue-engineered androgen secretory tissue implant

A technology of tissue engineering and secretory tissue, which is applied in the fields of medicine and biomedical engineering, and can solve the problems of inability to achieve therapeutic effects and short-term maintenance of hormone levels

Inactive Publication Date: 2011-02-16
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current domestic and foreign research on tissue engineering technology to construct androgen-secreting tissues shows that although tissue engineering methods are technically feasible to construct androgen-secreting tissues, the hormone levels of the constructed tissues are maintained for a short time after transplantation, and long-term stable therapeutic effects cannot be achieved.

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preparation example Construction

[0060] A preferred preparation method is to cut the testis into small pieces, digest it with 0.3% compound collagenase until all the tissue structures disappear and become muddy, filter through a Φ100 μm filter screen, centrifuge the filtrate, and get the precipitated cells to inoculate and cultivate, serum-free DMEM / F12 ( 1:1) After 24 hours of culture, all adherent cells were collected to obtain co-cultured testicular somatic cells.

[0061] A more preferred preparation method is to separate and cultivate LC and SC to obtain LC and SC respectively, both of which have a purity of 90-100%, and mix them according to a ratio of 1:0.01-30 to obtain co-cultured testicular somatic cells, a preferred The ratio of LC to SC is 1:0.1-1.

[0062] Testicular Somatic Cell Mass Uses

[0063] The testicular somatic cell population provided by the present invention is a new seed cell, which can be used to construct tissue-engineered androgen-secreting tissue, which is a complex of testicula...

Embodiment 1

[0089] Preparation of Testicular Somatic Cell Population I - Direct Adherence Co-culture Method

[0090] The rat testis tissue from which the buffy coat had been removed was digested with 0.25% collagenase NB4 (Serva, Germany) and the amount of tissue at a ratio of 1:1, 37°C, 100 times / min, shaking and digesting until the shape of the testis tissue completely disappeared and became muddy, adding 3 times the volume of PBS to stop the enzyme digestion, filter with a stainless steel filter with a pore size of 100um, centrifuge at 1500 rpm for 5 minutes, discard the supernatant, and inoculate the obtained cells in a culture dish, with DMEM / F12, 37 Cultivate under the conditions of ℃, 5% CO2 and saturated humidity, change the medium for the first time after 24 hours, continue to cultivate in the original medium and culture conditions until 72 hours, and the obtained cells are the testicular somatic cell population obtained by direct adherent co-culture.

[0091] Observation under a...

Embodiment 2

[0093] Preparation of testicular somatic cell population II-LC mixed with SC

[0094] (1) LC separation and culture

[0095] Rat testicular tissue from which the buffy coat had been removed was digested with 0.25% collagenase NB4 (Serva, Germany) and tissue volume at a ratio of 1:1 at 37°C and 100 times / min until the shape of the testicular tissue just disappeared. When the tissue is loose, but the seminiferous tubules are continuous without obvious breakage, add culture medium or PBS twice the volume of the digestion suspension to stop the digestion, filter through a Φ=30 μm stainless steel filter, and centrifuge the filtrate at 1500 rpm for 5 minutes. The resulting pellet was inoculated with 20 cm of cells per gram of testicular tissue 2 inoculation density. 37°C, 5% CO 2 , 95%O 2 Under the condition of 100% humidity, serum-free DMEM / F12 (1:1) culture solution was cultured for 2 hours, and the supernatant and unattached suspension cells were discarded.

[0096] The resu...

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Abstract

The invention discloses a tissue-engineered androgen secretion tissue graft, comprising (1) pharmaceutically accepted biomaterial and (2) a testis body cell group, wherein, the testis body cell group comprises separated and purified Leydig cells (LC) and mixture of LC and other testis body cells. After the androgen secretion tissue disclosed by the invention is transplanted in the body, androgen required by various physiological activities can be effectively secreted. The invention also provides the specific manufacturing method and purposes of the androgen secretion tissue.

Description

technical field [0001] The invention relates to the fields of medicine and biomedical engineering, and more specifically relates to a tissue-engineered androgen-secreting tissue constructed from testicular somatic cells. Background technique [0002] Androgen is a male characteristic endocrine hormone, which is mainly synthesized and secreted by Leydig cells (LC) in the testicular interstitium, and participates in the metabolic process of various tissues and cells in the body. Testicular absence or dysfunction caused by various reasons will cause low androgen levels, which will have an important impact on the physiology and psychology of men at different developmental stages, and eventually lead to a serious decline in the quality of life of patients. In recent years, with the accelerated pace of life, increased work pressure, and the increase in the aging population, the incidence of such diseases has gradually increased. Unfortunately, there is no safe and effective clini...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38
Inventor 王晓云周广东邢新
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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