CN103114072B - Method for largely extracting active epithelial cells - Google Patents
Method for largely extracting active epithelial cells Download PDFInfo
- Publication number
- CN103114072B CN103114072B CN201310016697.7A CN201310016697A CN103114072B CN 103114072 B CN103114072 B CN 103114072B CN 201310016697 A CN201310016697 A CN 201310016697A CN 103114072 B CN103114072 B CN 103114072B
- Authority
- CN
- China
- Prior art keywords
- active epithelial
- large amount
- active
- pancreatin
- cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 238000000034 method Methods 0.000 title claims abstract description 28
- 210000002919 epithelial cell Anatomy 0.000 title abstract description 11
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000000741 silica gel Substances 0.000 claims abstract description 13
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 13
- 210000001519 tissue Anatomy 0.000 claims abstract description 13
- 230000003472 neutralizing effect Effects 0.000 claims abstract description 5
- 238000004140 cleaning Methods 0.000 claims abstract description 4
- 210000003491 skin Anatomy 0.000 claims description 19
- 108010019160 Pancreatin Proteins 0.000 claims description 13
- 229940055695 pancreatin Drugs 0.000 claims description 13
- 238000000605 extraction Methods 0.000 claims description 12
- 229960001866 silicon dioxide Drugs 0.000 claims description 12
- 241001465754 Metazoa Species 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 210000000434 stratum corneum Anatomy 0.000 claims description 4
- 108010039627 Aprotinin Proteins 0.000 claims description 3
- 230000029087 digestion Effects 0.000 claims description 3
- 238000004321 preservation Methods 0.000 claims description 3
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims description 2
- 244000045232 Canavalia ensiformis Species 0.000 claims description 2
- 102000002322 Egg Proteins Human genes 0.000 claims description 2
- 108010000912 Egg Proteins Proteins 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 claims description 2
- 235000010617 Phaseolus lunatus Nutrition 0.000 claims description 2
- 229960004405 aprotinin Drugs 0.000 claims description 2
- 229940069765 bean extract Drugs 0.000 claims description 2
- 235000014103 egg white Nutrition 0.000 claims description 2
- 210000000969 egg white Anatomy 0.000 claims description 2
- 239000000284 extract Substances 0.000 claims description 2
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 claims description 2
- 239000007769 metal material Substances 0.000 claims description 2
- 244000005700 microbiome Species 0.000 claims description 2
- 239000004033 plastic Substances 0.000 claims description 2
- 229920003023 plastic Polymers 0.000 claims description 2
- 210000002966 serum Anatomy 0.000 claims description 2
- 230000009261 transgenic effect Effects 0.000 claims description 2
- -1 wooden Polymers 0.000 claims 2
- 238000009966 trimming Methods 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 17
- 210000000981 epithelium Anatomy 0.000 abstract description 9
- 108090000790 Enzymes Proteins 0.000 abstract description 7
- 102000004190 Enzymes Human genes 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 6
- 230000006378 damage Effects 0.000 abstract description 5
- 238000001976 enzyme digestion Methods 0.000 abstract 1
- 239000002245 particle Substances 0.000 abstract 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 3
- 229910052710 silicon Inorganic materials 0.000 description 3
- 239000010703 silicon Substances 0.000 description 3
- 230000007704 transition Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 206010058820 Acantholysis Diseases 0.000 description 1
- 101000984722 Bos taurus Pancreatic trypsin inhibitor Proteins 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 210000000270 basal cell Anatomy 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 210000002503 granulosa cell Anatomy 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000010297 mechanical methods and process Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 206010033675 panniculitis Diseases 0.000 description 1
- 238000009527 percussion Methods 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 210000000438 stratum basale Anatomy 0.000 description 1
- 210000000437 stratum spinosum Anatomy 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Images
Abstract
The invention discloses a method for largely extracting active epithelial cells and belongs to the technical field of biomedical science. The method comprises the following steps of: removing cuticles and partial granular layer cells by adopting a buffing method; removing epidermal skin grafts by a roller type dermatome; horizontally paving the removed epidermal skin grafts on a large silica gel plate; cutting the epidermal skin grafts into micro tissue blocks by a particle cutter; putting the micro tissue blocks to pancreatic enzyme for being digested for 1 minute to 10 minutes, so that the tissue blocks are loosened into cells or cell clusters; and neutralizing pancreatic enzyme by neutralizer and cleaning the neutralized pancreatic enzyme, and storing the cleaned pancreatic enzyme for future use. The method disclosed by the invention is adopted, so that not only 90% of removed skin is active epithelial tissues, but also the damages to the active epithelial tissues are greatly reduced. The micro epithelial tissues are digested to be active epithelial cells and cell clusters by the pancreatic enzyme, so that the damages caused by excessive enzyme digestion are avoided, and the activities of the active epithelial cells and cell clusters are improved.
Description
Technical field
The invention belongs to field of biomedicine technology, be specifically related to a kind of active epithelial method of a large amount of extraction.
Background technology
Skin is the organ of human body maximum, is also directly connected to people's image, beautiful and health except various physiological functions.Therefore, to skin epithelial research be engineering in medicine be also beautiful engineering.
As the outermost layer of skin, except palm sole of the foot position, epithelium is generally divided into four layers, is outside in followed successively by:
(1) deactivated stratum corneum, with age, sex, ethnic group and slightly different, is generally 5-10 layer, and palm sole of the foot position can thickly reach 40-50 layer;
(2) granular layer in active epithelial cell and dead keratinocyte transition period, general about layer 2-4;
(3) still possess the stratum spinosum epidermidis of division growth ability, by 4-8 layer spinose cell, formed;
(4) the most vigorous stratum basale of division growth ability, also referred to as malpighian layer, is comprised of the cylindric basal cell of individual layer.
Gather epithelial method in the past and be generally the method for getting holostrome or medium bed skin and then cutting with mechanical method or enzyme and removed various unwanted weave constructions, filter out activated epithelial cell (being basal layer cell or acantholysis cell), not only waste time and energy, exhaust the people and drain the treasury, and a large amount of active epithelial cells come to harm in gatherer process, affect its surviving rate, active epithelial number is declined greatly, get skin zone position simultaneously and also can leave scar in various degree, be unsuitable for epithelial commercialization and use.
Summary of the invention
In order effectively to solve the difficult problem existing in prior art, the invention provides a kind of active epithelial method of a large amount of extraction, described method comprises the steps:
The first step: first remove stratum corneum and part granular layer cell by the method for mill skin;
Second step: get razor graft with Humby knife;
The 3rd step: the razor graft taking off is laid on large silica-gel plate;
The 4th step: adopt particulate cutters that razor graft is cut into miniature tissue block;
The 5th step: miniature tissue block is put into pancreatin at 0-40 degree Celsius of digestion 1-10 minute, make to organize loose one-tenth cell or cell bulk;
The 6th step: in and pancreatin eccentric cleaning, 0-40 degree Celsius of preservation is stand-by.
Advantage of the present invention is:
1, first removed the Eponychium of non-activity and lacked on a small quantity active granulosa cell;
2, razor graft is got skin and has been reduced again subcutis, and making got skin more than 90% is active epithelium;
3, with the special sharp property of miniature dermatotome skin bit in activity is cut into rapidly to miniature epithelium, greatly reduced the damage of active epithelium;
4, with pancreatin, miniature epithelium is digested to active epithelial cell and cell mass, has avoided again transition enzyme to cut the injury causing, promoted the activity of active epithelial cell and cell mass;
5,, according to theory and practice research, the activity of epithelial cell group is far longer than single epithelial activity, has more business applying value.
Accompanying drawing explanation
Fig. 1 is the schema of the active epithelial method of a large amount of extraction of the present invention;
Fig. 2 a is the schematic cross-section of the particulate cutters that adopt in the present invention;
Fig. 2 b is the cutting knife design diagram of the particulate cutters that adopt in the present invention.
Embodiment
Below in conjunction with drawings and Examples, the present invention is described in detail.
The invention provides a kind of active epithelial method of a large amount of extraction, flow process, specifically comprises the steps: as shown in Figure 1
The first step: first remove gently stratum corneum and part granular layer cell by the method for mill skin, the skin of take exposes pink as degree; The method of described mill skin comprises the methods such as machinery, laser or chemistry mill skin.General 1～3 layer of the granular layer of removing.
Second step: get razor graft by required size with Humby knife; It is 0.15～0.25mm that described Humby knife is set thickness, the about 0.2mm of the thinnest thickness, and the thickness of the skin of getting is set as 0.15～0.25mm.
The 3rd step: the razor graft taking off is laid on large silica-gel plate.
Described large silica-gel plate size is about 10 * 10cm
2, in order to the skin graft that takes off of carrying, silica-gel plate can adopt the materials such as plastics, wooden or rubber, silica gel, and hardness is less than the hardness of metallic substance, but can carry again cutting and meet the easily requirement of cutting.
The 4th step: adopt particulate cutters that razor graft is cut into miniature tissue block.
As shown in Figure 2 a and 2 b, thickness is 0.5～2cm to described particulate cutters.Cut surface 2 and hammering face 1 two portions of comprising one-body molded processing, the cutting knife that described cut surface 2 is mesh design, cutting knife has the very sharp keen knife edge, is the blade of cutting knife, and as shown in Figure 1, cutting knife is designed to fine and closely woven latticed; Size 0.02～the 2mm of each grid, size is the smaller the better, is generally preferably 0.02～1mm.
Described hammering face 1 is non-cut surface, and cut surface is thicker blunt relatively, and for carrying percussion power, hammering face 1 is generally the blunt type square of 2 * 2cm.
During cutting, only the razor graft taking off need be laid on bulk silicon offset plate, on fritter silica-gel plate horizontal hammering face, cutting knife is aimed at the razor graft on the bulk silicon offset plate that will cut, and appropriate hammering can obtain miniature tissue block.As required, generally also optional majority particulate cutters simultaneously on the skin histology of horizontal on bulk silicon offset plate, evenly and fast knock fritter silica-gel plate to knock hammer, can obtain rapidly a large amount of miniature epithelium pieces, can be for directly or further.
The area size of described little silica-gel plate and described hammering face equal and opposite in direction.
The 5th step: miniature tissue block is put into pancreatin and digest 1～10 minute, make the loose one-tenth cell of described miniature tissue block or cell bulk.Must guard against transition digestion.
The 6th step: with also cleaning with pancreatin in equal-volume neutralizing agent, maximum obtains active epithelial cell and cell mass, and 0-40 degree Celsius of preservation is stand-by.
Described pancreatin can be the pancreatin that animal tissues extracts, and can be also transgenic plant, the restructuring pancreatin that zooblast and microorganism are extracted.
Described neutralizing agent is animal serum, or STI, or lima bean extract, or egg white, or BPTI(bovine pancreatic trypsin inhibitor), or aprotinin.
Claims (7)
1. the active epithelial method of a large amount of extraction, is characterized in that: comprise the steps:
The first step: first remove stratum corneum and part granular layer cell by the method for mill skin, remove 1～3 layer of appearance granular layer;
Second step: get razor graft with Humby knife; It is 0.15mm～0.25mm that described Humby knife is set thickness;
The 3rd step: the razor graft taking off is laid on large silica-gel plate;
The 4th step: adopt particulate cutters that razor graft is cut into miniature tissue block;
The 5th step: miniature tissue block is put into pancreatin at 0-40 degree Celsius of digestion 1-10 minute, so that organize loose one-tenth cell or cell bulk;
The 6th step: use in neutralizing agent and pancreatin eccentric cleaning, stand-by 0-40 degree Celsius of preservation.
2. the active epithelial method of a kind of a large amount of extractions according to claim 1, is characterized in that: the method for described mill skin comprises machinery, laser or chemistry mill skin.
3. the active epithelial method of a kind of a large amount of extractions according to claim 1, is characterized in that: described large silica-gel plate is of a size of 10 * 10cm
2, the razor graft taking off in order to carrying.
4. the active epithelial method of a kind of a large amount of extractions according to claim 1, it is characterized in that: the thickness of described particulate cutters is 0.5～2cm, cut surface and hammering face two portions of comprising one-body molded processing, the cutting knife that described cut surface is mesh design; The trimming size 0.02～2mm of each square node; Described hammering face is non-cut surface, the blunt type square that hammering face is 2 * 2cm.
5. the active epithelial method of a kind of a large amount of extractions according to claim 1, is characterized in that: silica-gel plate adopts plastics, wooden, rubber or silica gel, and hardness is less than the hardness of metallic substance.
6. the active epithelial method of a kind of a large amount of extractions according to claim 1, is characterized in that: pancreatin is the pancreatin that animal tissues extracts, or transgenic plant, the restructuring pancreatin that zooblast and microorganism are extracted.
7. the active epithelial method of a kind of a large amount of extractions according to claim 1, is characterized in that: described neutralizing agent is animal serum, or STI, or lima bean extract, or egg white, or uBPTI, or aprotinin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310016697.7A CN103114072B (en) | 2013-01-16 | 2013-01-16 | Method for largely extracting active epithelial cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310016697.7A CN103114072B (en) | 2013-01-16 | 2013-01-16 | Method for largely extracting active epithelial cells |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103114072A CN103114072A (en) | 2013-05-22 |
| CN103114072B true CN103114072B (en) | 2014-04-02 |
Family
ID=48412489
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310016697.7A Expired - Fee Related CN103114072B (en) | 2013-01-16 | 2013-01-16 | Method for largely extracting active epithelial cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103114072B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1865436A (en) * | 2006-03-27 | 2006-11-22 | 西北农林科技大学 | Method of separating derma epidermis stem cell |
| CN101765657A (en) * | 2007-11-09 | 2010-06-30 | Rnl生物技术株式会社 | Method for isolating and culturing adult stem cells derived from human amniotic epithelium |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0608403D0 (en) * | 2006-04-28 | 2006-06-07 | Univ Durham | Substrate for the routine growth of cultured cells in three dimensions |
-
2013
- 2013-01-16 CN CN201310016697.7A patent/CN103114072B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1865436A (en) * | 2006-03-27 | 2006-11-22 | 西北农林科技大学 | Method of separating derma epidermis stem cell |
| CN101765657A (en) * | 2007-11-09 | 2010-06-30 | Rnl生物技术株式会社 | Method for isolating and culturing adult stem cells derived from human amniotic epithelium |
Non-Patent Citations (6)
| Title |
|---|
| 3种方法联合运用提取人体脱落上皮细胞DNA;郭燕霞等;《法医学杂志》;20090225(第01期);全文 * |
| 夏荔.皮肤 人体第一道生命防线.《健康天地》.2004 |
| 孙秀柱等.鲁西黄牛皮肤成纤维细胞分离与培养的研究.《中国畜牧兽医》.2004,(第12期), |
| 皮肤,人体第一道生命防线;夏荔;《健康天地》;20040405(第07期);第39页 * |
| 郭燕霞等.3种方法联合运用提取人体脱落上皮细胞DNA.《法医学杂志》.2009,(第01期), |
| 鲁西黄牛皮肤成纤维细胞分离与培养的研究;孙秀柱等;《中国畜牧兽医》;20041220(第12期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103114072A (en) | 2013-05-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2020202310B2 (en) | Pixel array medical devices and methods | |
| EP2928395B1 (en) | Pixel array medical devices | |
| Nyame et al. | Clinical applications of skin substitutes | |
| Rose | The latest innovations in hair transplantation | |
| CN110312482B (en) | Pixel array medical system, device and method | |
| JP5153638B2 (en) | Method for delivering hair follicle progenitor cells to the skin | |
| US20200275945A1 (en) | Pixel array medical devices and methods | |
| CN103908424A (en) | Beautifying and nursing essence and preparation method thereof | |
| Rubis et al. | The use of split-thickness dermal grafts to resurface full thickness skin defects | |
| CN103114072B (en) | Method for largely extracting active epithelial cells | |
| WO2014117597A2 (en) | Rapid tissue engineering method for skin construction and repair of skin damage | |
| CN110891503A (en) | Pixel array medical system, device and method | |
| RU2405478C1 (en) | Method of autodermoplasty with split perforated transplants in case of burns | |
| CN101468213A (en) | Method for preparing irradiation crosslinking heterogeneous skin acellular matrix and products produced thereby | |
| KR102151501B1 (en) | Composition for improving hair and head skin | |
| US11490952B2 (en) | Pixel array medical devices and methods | |
| SE1650588A1 (en) | Multi-layer dermatome | |
| TW200516150A (en) | Method for preparing papilla cells-containing preparation, composition and method for hair follicle reconstitution, and animal with reconstituted hair follicle | |
| Bahadoran et al. | Classification of surgical therapies for vitiligo | |
| Imahara et al. | Skin grafts | |
| Mayhew | Creeping eruption caused by the larvae of the cattle hookworm Bunostomum phlebotomum | |
| KR102382212B1 (en) | Pixel array medical devices and methods | |
| Goreshi et al. | Tissue conserving technique for reconstruction of nasal lining defects | |
| Den Hollander et al. | Covering the massive burn: a case series and review of the literature | |
| CN209518904U (en) | A kind of crushing of human fat tissue is cut |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: BEJING BOSHIYUAN HAIR RESEARCH CENTER Free format text: FORMER OWNER: GUANGDONG ZHAOGANG BIOTECHNOLOGY SCIENCE + TECHNOLOGY CO., LTD. Effective date: 20140224 |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 523808 DONGGUAN, GUANGDONG PROVINCE TO: 100022 CHAOYANG, BEIJING |
|
| TA01 | Transfer of patent application right |
Effective date of registration: 20140224 Address after: No. 5 C 100022 Beijing city 100 Garden Room 206 Baiziwan Applicant after: GUANGDONG ZHAOGANG BIOLOGICAL SCIENCE & TECHNOLOGY CO., LTD. Address before: 523808, building 2, building 201, Songshan science and Technology Industrial Park, building 10, Songshan Lake, Guangdong, Dongguan Applicant before: Guangdong Zhaogang Biological Science & Technology Co., Ltd. |
|
| TA01 | Transfer of patent application right | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160217 Address after: 100023 Beijing city Chaoyang District boziwan 100 Park Building 4, block A 901 trial Patentee after: BEIJING BOSHIYUAN SCIENCE AND TECHNOLOGY DEVELOPMENT CO., LTD. Address before: No. 5 C 100022 Beijing city 100 Garden Room 206 Baiziwan Patentee before: GUANGDONG ZHAOGANG BIOLOGICAL SCIENCE & TECHNOLOGY CO., LTD. |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140402 Termination date: 20200116 |