CN102763642A - Cryoprotectant and method for cryopreserving placenta amnion and chorion - Google Patents
Cryoprotectant and method for cryopreserving placenta amnion and chorion Download PDFInfo
- Publication number
- CN102763642A CN102763642A CN2012102887063A CN201210288706A CN102763642A CN 102763642 A CN102763642 A CN 102763642A CN 2012102887063 A CN2012102887063 A CN 2012102887063A CN 201210288706 A CN201210288706 A CN 201210288706A CN 102763642 A CN102763642 A CN 102763642A
- Authority
- CN
- China
- Prior art keywords
- chorion
- amnion
- dextran
- dimethyl sulfoxide
- placenta
- 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.)
- Granted
Links
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to the technical field of tissue engineering and discloses a cryoprotectant and a method for cryopreserving placenta amnion and chorion. The cryoprotectant is composed of fetal calf serum, dimethyl sulfoxide and dextran 40, and the volume ratio of the fetal calf serum, the dimethyl sulfoxide and the dextran 40 is 7-9:1:1. On the basis of deep research of placenta amnion and chorion structures, the dimethyl sulfoxide, the dextran 40 and the fetal calf serum suitable for protecting placenta amnion stem cells and chorion stem cells are selected to form the cryoprotectant. The fetal calf serum, the dimethyl sulfoxide and the dextran 40 coordinate to protect activity of cells, the cells are prevented from forming ice crystal and being damaged, and the activity of recovered stem cells and the activity of newly prepared amnion chorion stem cells have no difference.
Description
Technical field
The present invention relates to the tissue engineering technique field, relate to a kind of frozen solution and freezing depositary's placenta amnion and chorial method specifically.
Background technology
Amnion and chorion are two chief components of people's placenta.Amnion is at the innermost layer of placenta, and film body is smooth, no blood vessel, nerve and lymph; Has certain elasticity; It contains and abundant has multipotency, can not repel anti-multipotential stem cell by induction of immunity after transplanting, and amniotic cell of human placenta also can be expressed the nerve cell specific mark, the secretory nerve trophic factors; Be used for nervous system disease, in addition people's placenta amnion can also be as the basilar memebrane of ophthalmologic operation; And chorion is the placenta main part, and is adjacent with amnion, contains the abundant stem cell with many differentiation potentials on the chorion equally, can be applicable to because in injuries of tissues and organs reparation that aging and pathology cause and the transplanting.
Utopian engineering tissue except needs possess very high biologically active, also needs to satisfy at any time and transplants the needs that wait operation, and therefore, the engineering tissue that as people's placenta amnion and chorion, contains multipotential stem cell just needs long preservation.It is the common method that biological tissue is preserved that cryogenic freezing is preserved, and it generally is to preserve at 0～-196 ℃.Although cryogenic freezing technology can make the engineering tissue long preservation, also brought the problem of freezing injury.In order to reduce the problem of freezing injury, when preserving cell and tissue, need add frozen solution, thereby alleviate or avoid cooling off, the damage of the ice crystal pair cell in the recovery process.
The patent of one Chinese patent application numbers 201010528721.1 provides a kind of " the frozen protection liquid of people's umbilical cord China's Tong Shi glue piece of tissue "; It is according to the characteristics of claimed mescenchymal stem cell; Disclose by the multiple one-tenth of permeability cryoprotector, impermeability cryoprotector and basal liquid or the like and be grouped into protection liquid, the degree of impairment that the reply mescenchymal stem cell possibly occur in freezing preservation.But, do not have as yet at present to people's placenta amnion and chorial cryogenic freezing preservation technology report.Therefore, a kind of freezing depositary's placenta amnion and chorial technology are provided, can improve amnion and chorial preservation quality undoubtedly, guarantee the effect of amnion and the clinical practice of chorion stem cell.
Summary of the invention
In view of this, the object of the present invention is to provide a kind of frozen solution and freezing depositary's placenta amnion and chorial method, make said method can effectively protect people's placenta amnion and chorion, make its recovery after the stem cell activity reach more than 90%.
To achieve these goals, the present invention provides following technical scheme:
A kind of frozen solution is made up of hyclone, dimethyl sulfoxide (DMSO) and Dextran 40, and the volume ratio of said hyclone, dimethyl sulfoxide (DMSO) and Dextran 40 is 7-9:1:1.
Wherein, as preferably, the volume ratio of said hyclone, dimethyl sulfoxide (DMSO) and Dextran 40 is 8:1:1.
Dimethyl sulfoxide (DMSO) (DMSO) is a kind of important osmosis type cell-protecting.When profound hypothermia (subzero 196 degree) preservation cell, in the frozen process, can prevent the damage that the formation of intracellular fluid ice crystal, osmotic pressure change, cell structure disorder etc. cause.DMSO can get in the cell by the quick penetration cell membrane, reduces freezing point, delays frozen process, improves intracellular ion concentration simultaneously, reduces the formation of ice crystal in the cell, thereby reduces cellular damage.And Dextran 40 belongs to the impermeability antifreeze, can be water-soluble, but can not get into cell, and make solution be supercooled state, can under specified temp, reduce solute concentration, thereby play a protective role.Because its molecular weight is big, molecular concentration is low, and is very little to the active function of solvent, can reduce low molecule solute (electrolyte) concentration in the solution, alleviates the salt damage, and the slow freezing effect is better.Freeze-stored cell is a process slowly, and hyclone provides necessary nutrition to freeze-stored cell, and contains the rich in protein pair cell in the serum protective effect is also arranged.
Hyclone, Dextran 40 and DMSO according to mixed according to the invention as people's placenta amnion and chorion the protection liquid when frozen, Dextran 40 is the extracellular protective agent, DMSO has high osmosis; It is protectant in the cell; Hyclone is a nutriment, and the three is adapted to the characteristic of amnion stem cell and chorion stem cell, the coordinating protection cell activity; Prevent that cell from forming ice crystal in frozen and recovery, and then influence cell activity.Therefore, institute of the present invention book frozen solution can be applied in cryogenic freezing depositary placenta amnion and the chorion.
In addition; The present invention also provides a kind of freezing depositary's placenta amnion and chorial method; With hyclone: dimethyl sulfoxide (DMSO): Dextran 40 is 7-9:1: 1 volume ratio weighing hyclone, dimethyl sulfoxide (DMSO) and Dextran 40, be mixed with frozen solution, and people's placenta amnion and chorion to be stored are immersed in the frozen solution respectively; Be cooled to-100 ℃ then, be transferred in the liquid nitrogen at last and preserve.
As preferably, the volume ratio of said hyclone, dimethyl sulfoxide (DMSO) and Dextran 40 is 8:1:1,
Said cooling operation is preferably:
Earlier reduce to-40 ℃, and then lower the temperature with the speed of 5 ℃/min with the speed of 1 ℃/min.
To recover according to the people's placenta amnion and the chorion of the frozen mistake of the method for the invention; Separation, culturing stem cells are carried out the activity contrast with the stem cell that utilizes the new people's placenta amnion gathered and chorion to prepare then, and both activities is all 90%; No significant difference; Shown that frozen solution according to the invention and method can extremely effectively protect amnion and chorionic cells active, cell does not almost receive freezing injury, with freshly prepd amnion and chorion stem cell indifference.
Can know by above technical scheme; The present invention is furtheing investigate on the structural basis of people's placenta amnion and chorion; Selection is suitable for protecting dimethyl sulfoxide (DMSO), Dextran 40 and the hyclone of people's placenta amnion stem cell and chorion stem cell to form protection liquid; Three's coordinating protection cell activity prevents that cell from forming ice crystal and sustaining damage, and has guaranteed the stem cell after the recovery active and freshly prepd amnion and the active indifference of chorion stem cell.
Embodiment
The embodiment of the invention discloses a kind of frozen solution and freezing depositary's placenta amnion and chorial method.Those skilled in the art can use for reference this paper content, suitably improve technological parameter and realize.Special needs to be pointed out is that all similarly replace and change apparent to those skilled in the art, they all are regarded as and are included in the present invention.Product of the present invention and method are described through preferred embodiment; The related personnel obviously can be in not breaking away from content of the present invention, spirit and scope to product as herein described and method is changed or suitably change and combination, realize and use technology of the present invention.
In order further to understand the present invention, a kind of frozen solution provided by the invention and freezing depositary's placenta amnion and chorial method are elaborated below in conjunction with embodiment.
Embodiment 1: utilize frozen people's placenta amnion of the method for the invention and chorion
1, frozen method
Choose the placenta that infectious diseases such as hepatitis, syphilis, AIDS detect feminine gender and do not have obstetric complication, warp gets puerpera's informed consent and signs Informed Consent Form before the art.Prepare aseptic polystyrene liquid storage bottle before the art as transporting bottle, in adorn commercially available cell culture fluid, gather back 4 ℃ of conditions and deliver to frozen place in following 12 hours.
Under the gnotobasis, with pincers gently separate amnion and chorion, operating scissors clip amnion and chorion.Wash amnion and chorion respectively 2-3 time with physiological saline, remove dirts such as surperficial clot after, amnion and chorion are cut into about 1cm respectively in culture dish
2Size.
With hyclone: dimethyl sulfoxide (DMSO): Dextran 40 is volume ratio weighing hyclone, dimethyl sulfoxide (DMSO) and the Dextran 40 of 8:1:1; Be mixed with frozen solution; People's placenta amnion to be stored and chorion are immersed in respectively in the frozen solution, divide in the frozen pipe of packing into again, tighten the pipe lid.Frozen pipe is put into the programmed cooling appearance, reduce to-40 ℃ with the speed of 1 ℃/min earlier, and then be cooled to-100 ℃, at last frozen pipe is changed in the liquid nitrogen over to-196 ℃ of long preservation with the speed of 5 ℃/min.
2, active contrast
Again get fresh human placenta and separate amnion and chorion according to collection standard in 1, wash amnion and chorion respectively 2-3 time with physiological saline, remove dirts such as surperficial clot after, amnion and chorion are cut into about 1cm respectively in culture dish
2Size places centrifuge tube, adds 0.2% II Collagen Type VI enzyme, 37 ℃ of water-bath digestion 40min.Add physiological saline toward centrifuge tube, 300 eye mesh screens filter; The centrifugal 5min of filtrating 300G washs 2-3 time.The perfect medium re-suspended cell is inoculated in the blake bottle then, in 37 ℃, 5%CO
2Cultivate.Approximately after 12-24 hour, observation of cell is adherent can to carry out changing first time liquid, after this whenever changes liquid at a distance from 3 days, treats promptly to obtain stem cell after cell grows to fusion, and detection stem cell activity reaches more than 90%.
Amnion frozen in 1 and chorion are taken out, put into immediately in 37 ℃ of water-baths, and rock rapidly, treat to take out frozen pipe immediately after the cryopreserving liquid dissolving.After the frozen tube-surface sterilization, in superclean bench, open, amnion and chorion suspension are transferred in the centrifuge tube, add precooling physiological saline, the centrifugal 5min of 300G washs 2-3 time.The method for preparing stem cell according to the amnion and the chorion of aforementioned new collection placenta afterwards, preparation recovery back amnion stem cell and chorion stem cell, detection stem cell activity reaches more than 90% equally.
The result shows, amnion and chorion stem cell after frozen almost do not receive freezing injury, and with freshly prepd amnion and chorion stem cell indifference, cytoactive is all more than 90%, and both cells grow to, and to merge the used time also identical.
Embodiment 2: utilize frozen people's placenta amnion of the method for the invention and chorion
1, frozen method
Choose the placenta that infectious diseases such as hepatitis, syphilis, AIDS detect feminine gender and do not have obstetric complication, warp gets puerpera's informed consent and signs Informed Consent Form before the art.Prepare aseptic polystyrene liquid storage bottle before the art as transporting bottle, in adorn commercially available cell culture fluid, gather back 4 ℃ of conditions and deliver to frozen place in following 12 hours.
Under the gnotobasis, with pincers gently separate amnion and chorion, operating scissors clip amnion and chorion.Wash amnion and chorion respectively 2-3 time with physiological saline, remove dirts such as surperficial clot after, amnion and chorion are cut into about 1cm respectively in culture dish
2Size.
With hyclone: dimethyl sulfoxide (DMSO): Dextran 40 is volume ratio weighing hyclone, dimethyl sulfoxide (DMSO) and the Dextran 40 of 7:1:1; Be mixed with frozen solution; People's placenta amnion to be stored and chorion are immersed in respectively in the frozen solution, divide in the frozen pipe of packing into again, tighten the pipe lid.Frozen pipe is put into the programmed cooling appearance, reduce to-40 ℃ with the speed of 1 ℃/min earlier, and then be cooled to-100 ℃, at last frozen pipe is changed in the liquid nitrogen over to-196 ℃ of long preservation with the speed of 5 ℃/min.
2, active contrast
Again get fresh human placenta and separate amnion and chorion according to collection standard in 1, wash amnion and chorion respectively 2-3 time with physiological saline, remove dirts such as surperficial clot after, amnion and chorion are cut into about 1cm respectively in culture dish
2Size places centrifuge tube, adds 0.2% II Collagen Type VI enzyme, 37 ℃ of water-bath digestion 40min.Add physiological saline toward centrifuge tube, 300 eye mesh screens filter; The centrifugal 5min of filtrating 300G washs 2-3 time.The perfect medium re-suspended cell is inoculated in the blake bottle then, in 37 ℃, 5%CO
2Cultivate.Approximately after 12-24 hour, observation of cell is adherent can to carry out changing first time liquid, after this whenever changes liquid at a distance from 3 days, treats promptly to obtain stem cell after cell grows to fusion, and detection stem cell activity reaches more than 90%.
Amnion frozen in 1 and chorion are taken out, put into immediately in 37 ℃ of water-baths, and rock rapidly, treat to take out frozen pipe immediately after the cryopreserving liquid dissolving.After the frozen tube-surface sterilization, in superclean bench, open, amnion and chorion suspension are transferred in the centrifuge tube, add precooling physiological saline, the centrifugal 5min of 300G washs 2-3 time.The method for preparing stem cell according to the amnion and the chorion of aforementioned new collection placenta afterwards, preparation recovery back amnion stem cell and chorion stem cell, detection stem cell activity reaches more than 90% equally.
The result shows, amnion and chorion stem cell after frozen almost do not receive freezing injury, and with freshly prepd amnion and chorion stem cell indifference, cytoactive is all more than 90%, and both cells grow to, and to merge the used time also identical.
Embodiment 3: utilize frozen people's placenta amnion of the method for the invention and chorion
1, frozen method
Choose the placenta that infectious diseases such as hepatitis, syphilis, AIDS detect feminine gender and do not have obstetric complication, warp gets puerpera's informed consent and signs Informed Consent Form before the art.Prepare aseptic polystyrene liquid storage bottle before the art as transporting bottle, in adorn commercially available cell culture fluid, gather back 4 ℃ of conditions and deliver to frozen place in following 12 hours.
Under the gnotobasis, with pincers gently separate amnion and chorion, operating scissors clip amnion and chorion.Wash amnion and chorion respectively 2-3 time with physiological saline, remove dirts such as surperficial clot after, amnion and chorion are cut into about 1cm respectively in culture dish
2Size.
With hyclone: dimethyl sulfoxide (DMSO): Dextran 40 is volume ratio weighing hyclone, dimethyl sulfoxide (DMSO) and the Dextran 40 of 9:1:1; Be mixed with frozen solution; People's placenta amnion to be stored and chorion are immersed in respectively in the frozen solution, divide in the frozen pipe of packing into again, tighten the pipe lid.Frozen pipe is put into the programmed cooling appearance, reduce to-40 ℃ with the speed of 1 ℃/min earlier, and then be cooled to-100 ℃, at last frozen pipe is changed in the liquid nitrogen over to-196 ℃ of long preservation with the speed of 5 ℃/min.
2, active contrast
Again get fresh human placenta and separate amnion and chorion according to collection standard in 1, wash amnion and chorion respectively 2-3 time with physiological saline, remove dirts such as surperficial clot after, amnion and chorion are cut into about 1cm respectively in culture dish
2Size places centrifuge tube, adds 0.2% II Collagen Type VI enzyme, 37 ℃ of water-bath digestion 40min.Add physiological saline toward centrifuge tube, 300 eye mesh screens filter; The centrifugal 5min of filtrating 300G washs 2-3 time.The perfect medium re-suspended cell is inoculated in the blake bottle then, in 37 ℃, 5%CO
2Cultivate.Approximately after 12-24 hour, observation of cell is adherent can to carry out changing first time liquid, after this whenever changes liquid at a distance from 3 days, treats promptly to obtain stem cell after cell grows to fusion, and detection stem cell activity reaches more than 90%.
Amnion frozen in 1 and chorion are taken out, put into immediately in 37 ℃ of water-baths, and rock rapidly, treat to take out frozen pipe immediately after the cryopreserving liquid dissolving.After the frozen tube-surface sterilization, in superclean bench, open, amnion and chorion suspension are transferred in the centrifuge tube, add precooling physiological saline, the centrifugal 5min of 300G washs 2-3 time.The method for preparing stem cell according to the amnion and the chorion of aforementioned new collection placenta afterwards, preparation recovery back amnion stem cell and chorion stem cell, detection stem cell activity reaches more than 90% equally.
The result shows, amnion and chorion stem cell after frozen almost do not receive freezing injury, and with freshly prepd amnion and chorion stem cell indifference, cytoactive is all more than 90%, and both cells grow to, and to merge the used time also identical.
The explanation of above embodiment just is used for helping to understand method of the present invention and core concept thereof.Should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention, can also carry out some improvement and modification to the present invention, these improvement and modification also fall in the protection domain of claim of the present invention.
Claims (6)
1. a frozen solution is characterized in that, is made up of hyclone, dimethyl sulfoxide (DMSO) and Dextran 40, and the volume ratio of said hyclone, dimethyl sulfoxide (DMSO) and Dextran 40 is 7-9:1:1.
2. according to the said frozen solution of claim 1, it is characterized in that the volume ratio of said hyclone, dimethyl sulfoxide (DMSO) and Dextran 40 is 8:1: 1.
3. the application of the said frozen solution of claim 1 in cryogenic freezing depositary placenta amnion and chorion.
4. freezing depositary's placenta amnion and chorial method; It is characterized in that; With hyclone: dimethyl sulfoxide (DMSO): Dextran 40 is volume ratio weighing hyclone, dimethyl sulfoxide (DMSO) and the Dextran 40 of 7-9:1:1, is mixed with frozen solution, and people's placenta amnion and chorion to be stored are immersed in the frozen solution respectively; Be cooled to-100 ℃ then, be transferred in the liquid nitrogen at last and preserve.
5. according to the said method of claim 4, it is characterized in that the volume ratio of said hyclone, dimethyl sulfoxide (DMSO) and Dextran 40 is 8:1:1.
6. according to the said method of claim 4, it is characterized in that said cooling is specially:
Earlier reduce to-40 ℃, and then lower the temperature with the speed of 5 ℃/min with the speed of 1 ℃/min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210288706.3A CN102763642B (en) | 2012-08-14 | 2012-08-14 | Cryoprotectant and method for cryopreserving placenta amnion and chorion |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210288706.3A CN102763642B (en) | 2012-08-14 | 2012-08-14 | Cryoprotectant and method for cryopreserving placenta amnion and chorion |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102763642A true CN102763642A (en) | 2012-11-07 |
| CN102763642B CN102763642B (en) | 2014-04-02 |
Family
ID=47091407
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210288706.3A Active CN102763642B (en) | 2012-08-14 | 2012-08-14 | Cryoprotectant and method for cryopreserving placenta amnion and chorion |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102763642B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105165803A (en) * | 2015-10-28 | 2015-12-23 | 广州赛莱拉干细胞科技股份有限公司 | Cryopreservation protection solution and cryopreservation method for placenta amniotic mesenchymal stem cells |
| CN108029679A (en) * | 2018-01-29 | 2018-05-15 | 山东省齐鲁细胞治疗工程技术有限公司 | A kind of frozen stock solution for being used to freeze mononuclearcell |
| CN108077243A (en) * | 2018-01-24 | 2018-05-29 | 北京臻溪谷医学研究中心（有限合伙） | A kind of freezen protective Human plactnta amnion and chorial protection liquid and preparation method thereof and application method |
| CN108812643A (en) * | 2018-07-18 | 2018-11-16 | 银丰生物工程集团有限公司 | Human placenia membrane tissue prepares cryopreservation methods and application |
| CN108812640A (en) * | 2018-07-18 | 2018-11-16 | 银丰生物工程集团有限公司 | Human plactnta amnion, decidua tissue prepare cryopreservation methods and application |
| CN109392891A (en) * | 2018-10-26 | 2019-03-01 | 银丰生物工程集团有限公司 | A kind of methods and applications freezing human umbilical tissue according to layer of structure system |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1728946A (en) * | 2002-07-19 | 2006-02-01 | 维斯塔治疗公司 | Method of obtaining viable human liver cells, including hepatic stem/progenitor cells |
| CN101257861A (en) * | 2005-06-21 | 2008-09-03 | 渗透治疗有限公司 | Methods and compositions for enhancing vascular access |
-
2012
- 2012-08-14 CN CN201210288706.3A patent/CN102763642B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1728946A (en) * | 2002-07-19 | 2006-02-01 | 维斯塔治疗公司 | Method of obtaining viable human liver cells, including hepatic stem/progenitor cells |
| CN101257861A (en) * | 2005-06-21 | 2008-09-03 | 渗透治疗有限公司 | Methods and compositions for enhancing vascular access |
Non-Patent Citations (1)
| Title |
|---|
| 李远等: "不同冻存液对人羊膜间充质干细胞-80℃冻存与复苏效果的影响", 《郑州大学学报（医学版）》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105165803A (en) * | 2015-10-28 | 2015-12-23 | 广州赛莱拉干细胞科技股份有限公司 | Cryopreservation protection solution and cryopreservation method for placenta amniotic mesenchymal stem cells |
| CN108077243A (en) * | 2018-01-24 | 2018-05-29 | 北京臻溪谷医学研究中心（有限合伙） | A kind of freezen protective Human plactnta amnion and chorial protection liquid and preparation method thereof and application method |
| CN108029679A (en) * | 2018-01-29 | 2018-05-15 | 山东省齐鲁细胞治疗工程技术有限公司 | A kind of frozen stock solution for being used to freeze mononuclearcell |
| CN108812643A (en) * | 2018-07-18 | 2018-11-16 | 银丰生物工程集团有限公司 | Human placenia membrane tissue prepares cryopreservation methods and application |
| CN108812640A (en) * | 2018-07-18 | 2018-11-16 | 银丰生物工程集团有限公司 | Human plactnta amnion, decidua tissue prepare cryopreservation methods and application |
| CN108812640B (en) * | 2018-07-18 | 2021-11-26 | 银丰生物工程集团有限公司 | Preparation and cryopreservation method and application of human placental amniotic membrane and decidua tissue |
| CN109392891A (en) * | 2018-10-26 | 2019-03-01 | 银丰生物工程集团有限公司 | A kind of methods and applications freezing human umbilical tissue according to layer of structure system |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102763642B (en) | 2014-04-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102763642B (en) | Cryoprotectant and method for cryopreserving placenta amnion and chorion | |
| CN108617638B (en) | Tissue and/or cell cryopreservation protective solution and preparation and application thereof | |
| CN104145943B (en) | A kind of frozen protection liquid of people's umbilical cord China Tong Shi glue tissue and preparation and application thereof | |
| CN101974484B (en) | Method for preparing human umbilical cord mesenchymal stem cells | |
| CN108207930A (en) | A kind of cocktail type cryoprotector and its application | |
| CN104472473B (en) | Umbilical cord tissue block cryopreservation methods | |
| CN102428911A (en) | Cryogenic storing protection liquid for human umbilical cord Wharton's jelly tissue block | |
| WO2009120996A1 (en) | Mehtod, system, and apparatus for hypothermic collection, storage, transport and banking of birth tissue | |
| CN110432259A (en) | A kind of frozen solution and the cells frozen storing liquid containing it and its application in cell cryopreservation | |
| CN108812643B (en) | Preparation and cryopreservation method and application of human placental chorionic villus tissue | |
| CN102428910A (en) | Protecting liquid for freezing and storing human umbilical cord Wharton jelly tissue block | |
| CN105211052B (en) | Frozen stock solution of cultured NKT cells and preparation method thereof | |
| CN107306940A (en) | The spermary Cryoprotectant and the preparation method of spermatogonium of a kind of seawater fish | |
| CN109699634A (en) | A kind of Cryopreservation and method for resuscitation of mescenchymal stem cell | |
| CN107494521B (en) | Cells frozen storing liquid and cell freezing method | |
| CN113925049B (en) | Cell preservation solution for maintaining cell activity and preparation method and application thereof | |
| CN106942200A (en) | One kind freezes protection liquid and its application | |
| CN113854280B (en) | Low-temperature preservation solution and preparation method and application thereof | |
| WO2001070021A1 (en) | Method of preserving tissue equivalent and tissue equivalent preserved in frozen state | |
| CN108812642B (en) | Systematic method for preparing placenta tissue according to structural hierarchy and cryopreserving and application | |
| CN107711823A (en) | The cells frozen storing liquid and its application that a kind of normal temperature preserves | |
| CN109042624B (en) | Preparation and cryopreservation method and application of human placental subpial large blood vessel tissue | |
| CN113041207A (en) | Preparation and recovery method of lyophilized powder of human mesenchymal stem cell culture supernatant | |
| CN107372469A (en) | The frozen stock solution and cryopreservation methods of a kind of endothelial progenitor cells | |
| CN112293407A (en) | Method for programmed cryopreservation of ovarian tissues |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: ZHENGZHOU YINGKE SAIER BIOLOGICAL ENGINEERING CO., Free format text: FORMER NAME: ZHENGZHOU CELLINCLONE STEM CELL TECHNOLOGY CO., LTD. |
|
| CP03 | Change of name, title or address |
Address after: 450000, No. 406, No. 2, South Building, 11 dragon road, hi tech District, Henan, Zhengzhou Patentee after: Zhengzhou Intco Purcell Biological Engineering Co. Ltd. Address before: 450000, South Building, No. 2, Pei Chun Road, 11 hi tech Road, Zhengzhou, Henan Province, 406 Patentee before: Zhengzhou Saiyingke Stem Cell Technology Co.,Ltd. |
|
| ASS | Succession or assignment of patent right |
Owner name: BOYA STEM CELLS TECHNOLOGY CO., LTD. Free format text: FORMER OWNER: ZHENGZHOU YINGKE SAIER BIOLOGICAL ENGINEERING CO., LTD. Effective date: 20140519 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 450000 ZHENGZHOU, HENAN PROVINCE TO: 214092 WUXI, JIANGSU PROVINCE |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20140519 Address after: Ma Shan Mei Binhu District 214092 in Jiangsu province Wuxi City Road No. 88 beam Patentee after: Boya Stem Cell Technology Co., Ltd. Address before: 450000, No. 406, No. 2, South Building, 11 dragon road, hi tech District, Henan, Zhengzhou Patentee before: Zhengzhou Intco Purcell Biological Engineering Co. Ltd. |