CN107530370A - 用于培养哺乳动物细胞的微载体、基质和支架及其制造方法 - Google Patents
用于培养哺乳动物细胞的微载体、基质和支架及其制造方法 Download PDFInfo
- Publication number
- CN107530370A CN107530370A CN201680017235.9A CN201680017235A CN107530370A CN 107530370 A CN107530370 A CN 107530370A CN 201680017235 A CN201680017235 A CN 201680017235A CN 107530370 A CN107530370 A CN 107530370A
- Authority
- CN
- China
- Prior art keywords
- copolymer
- polysaccharide
- polyamines
- selective oxidation
- glycoprotein
- 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
- 238000000034 method Methods 0.000 title claims abstract description 56
- 238000004519 manufacturing process Methods 0.000 title claims description 18
- 210000004962 mammalian cell Anatomy 0.000 title abstract description 29
- 239000011159 matrix material Substances 0.000 title abstract description 28
- 229920000768 polyamine Polymers 0.000 claims abstract description 102
- 229920001577 copolymer Polymers 0.000 claims abstract description 91
- 150000004676 glycans Chemical class 0.000 claims abstract description 65
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 56
- 239000005017 polysaccharide Substances 0.000 claims abstract description 56
- 239000002245 particle Substances 0.000 claims abstract description 49
- 239000008188 pellet Substances 0.000 claims abstract description 9
- 238000007254 oxidation reaction Methods 0.000 claims description 103
- 210000004027 cell Anatomy 0.000 claims description 101
- 230000003647 oxidation Effects 0.000 claims description 97
- 229920002678 cellulose Polymers 0.000 claims description 87
- 239000001913 cellulose Substances 0.000 claims description 84
- 229920000642 polymer Polymers 0.000 claims description 65
- 229920002873 Polyethylenimine Polymers 0.000 claims description 60
- 102000001621 Mucoproteins Human genes 0.000 claims description 42
- 108010093825 Mucoproteins Proteins 0.000 claims description 42
- 239000000463 material Substances 0.000 claims description 34
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 31
- 125000002091 cationic group Chemical group 0.000 claims description 24
- 239000000203 mixture Substances 0.000 claims description 23
- 150000001768 cations Chemical class 0.000 claims description 21
- 102000003886 Glycoproteins Human genes 0.000 claims description 19
- 108090000288 Glycoproteins Proteins 0.000 claims description 19
- 108090000623 proteins and genes Proteins 0.000 claims description 16
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 15
- 125000000524 functional group Chemical group 0.000 claims description 15
- 229910052757 nitrogen Inorganic materials 0.000 claims description 15
- 150000001299 aldehydes Chemical group 0.000 claims description 14
- 150000001412 amines Chemical class 0.000 claims description 14
- 102000004169 proteins and genes Human genes 0.000 claims description 14
- 229920000083 poly(allylamine) Polymers 0.000 claims description 13
- 125000003368 amide group Chemical group 0.000 claims description 11
- 238000007334 copolymerization reaction Methods 0.000 claims description 11
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 claims description 11
- 150000004804 polysaccharides Polymers 0.000 claims description 11
- 229920001661 Chitosan Polymers 0.000 claims description 10
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 9
- 239000000835 fiber Substances 0.000 claims description 9
- 229920002101 Chitin Polymers 0.000 claims description 8
- 229920002527 Glycogen Polymers 0.000 claims description 8
- 229940096919 glycogen Drugs 0.000 claims description 8
- 229920001503 Glucan Polymers 0.000 claims description 7
- 229920002472 Starch Polymers 0.000 claims description 7
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 7
- 239000008107 starch Substances 0.000 claims description 7
- 235000019698 starch Nutrition 0.000 claims description 7
- 229920001184 polypeptide Polymers 0.000 claims description 4
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 4
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 4
- AGGKEGLBGGJEBZ-UHFFFAOYSA-N tetramethylenedisulfotetramine Chemical compound C1N(S2(=O)=O)CN3S(=O)(=O)N1CN2C3 AGGKEGLBGGJEBZ-UHFFFAOYSA-N 0.000 claims description 4
- 229920001400 block copolymer Polymers 0.000 claims description 3
- 150000002466 imines Chemical class 0.000 claims description 3
- 230000005588 protonation Effects 0.000 claims description 3
- 210000000717 sertoli cell Anatomy 0.000 claims description 2
- 229920000856 Amylose Polymers 0.000 claims 3
- 229920000333 poly(propyleneimine) Polymers 0.000 claims 2
- 241001313855 Bletilla Species 0.000 claims 1
- 230000012010 growth Effects 0.000 abstract description 35
- 235000010980 cellulose Nutrition 0.000 description 84
- 239000000243 solution Substances 0.000 description 51
- 238000006243 chemical reaction Methods 0.000 description 29
- 238000001556 precipitation Methods 0.000 description 27
- 230000010261 cell growth Effects 0.000 description 23
- 238000003756 stirring Methods 0.000 description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 20
- 239000001963 growth medium Substances 0.000 description 17
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 17
- 239000006228 supernatant Substances 0.000 description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 15
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 14
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 14
- 238000004113 cell culture Methods 0.000 description 14
- 238000000576 coating method Methods 0.000 description 14
- 239000000047 product Substances 0.000 description 14
- 239000003643 water by type Substances 0.000 description 14
- 239000011248 coating agent Substances 0.000 description 13
- 238000004132 cross linking Methods 0.000 description 13
- 230000005484 gravity Effects 0.000 description 13
- 239000005090 green fluorescent protein Substances 0.000 description 13
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 11
- 238000013459 approach Methods 0.000 description 11
- 239000000725 suspension Substances 0.000 description 11
- -1 aldehyde radical Chemical class 0.000 description 10
- 229920006317 cationic polymer Polymers 0.000 description 9
- 239000003153 chemical reaction reagent Substances 0.000 description 9
- 239000004744 fabric Substances 0.000 description 9
- 230000014509 gene expression Effects 0.000 description 9
- 230000002018 overexpression Effects 0.000 description 9
- 229920000729 poly(L-lysine) polymer Polymers 0.000 description 9
- 235000018102 proteins Nutrition 0.000 description 9
- 238000006722 reduction reaction Methods 0.000 description 9
- 238000005406 washing Methods 0.000 description 9
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 8
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 7
- 238000013019 agitation Methods 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 230000009467 reduction Effects 0.000 description 7
- 239000011734 sodium Substances 0.000 description 7
- 229910052708 sodium Inorganic materials 0.000 description 7
- JQWHASGSAFIOCM-UHFFFAOYSA-M sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 description 7
- PFNFFQXMRSDOHW-UHFFFAOYSA-N spermine Chemical compound NCCCNCCCCNCCCN PFNFFQXMRSDOHW-UHFFFAOYSA-N 0.000 description 7
- DLFVBJFMPXGRIB-UHFFFAOYSA-N Acetamide Chemical compound CC(N)=O DLFVBJFMPXGRIB-UHFFFAOYSA-N 0.000 description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 6
- 239000008367 deionised water Substances 0.000 description 6
- 229910021641 deionized water Inorganic materials 0.000 description 6
- 230000004907 flux Effects 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 238000006116 polymerization reaction Methods 0.000 description 6
- 150000003141 primary amines Chemical class 0.000 description 6
- KIDHWZJUCRJVML-UHFFFAOYSA-N putrescine Chemical compound NCCCCN KIDHWZJUCRJVML-UHFFFAOYSA-N 0.000 description 6
- 239000002356 single layer Substances 0.000 description 6
- 239000000758 substrate Substances 0.000 description 6
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 5
- 206010052428 Wound Diseases 0.000 description 5
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 5
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 5
- 239000008187 granular material Substances 0.000 description 5
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 5
- 239000008108 microcrystalline cellulose Substances 0.000 description 5
- 229940016286 microcrystalline cellulose Drugs 0.000 description 5
- 230000000269 nucleophilic effect Effects 0.000 description 5
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 5
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- VHRGRCVQAFMJIZ-UHFFFAOYSA-N cadaverine Chemical compound NCCCCCN VHRGRCVQAFMJIZ-UHFFFAOYSA-N 0.000 description 4
- 150000001720 carbohydrates Chemical class 0.000 description 4
- 230000007541 cellular toxicity Effects 0.000 description 4
- 239000007795 chemical reaction product Substances 0.000 description 4
- 229920000359 diblock copolymer Polymers 0.000 description 4
- 235000013601 eggs Nutrition 0.000 description 4
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 4
- 230000007773 growth pattern Effects 0.000 description 4
- 210000005260 human cell Anatomy 0.000 description 4
- 239000007800 oxidant agent Substances 0.000 description 4
- 230000001590 oxidative effect Effects 0.000 description 4
- 239000000123 paper Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 239000001632 sodium acetate Substances 0.000 description 4
- 235000017281 sodium acetate Nutrition 0.000 description 4
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 4
- 235000017557 sodium bicarbonate Nutrition 0.000 description 4
- 229910000033 sodium borohydride Inorganic materials 0.000 description 4
- 239000012279 sodium borohydride Substances 0.000 description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L sodium carbonate Substances [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 229910000029 sodium carbonate Inorganic materials 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- XFNJVJPLKCPIBV-UHFFFAOYSA-N trimethylenediamine Chemical class NCCCN XFNJVJPLKCPIBV-UHFFFAOYSA-N 0.000 description 4
- 108010010803 Gelatin Proteins 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- 239000004793 Polystyrene Substances 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- 125000003172 aldehyde group Chemical group 0.000 description 3
- 125000003277 amino group Chemical group 0.000 description 3
- 229920003118 cationic copolymer Polymers 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 229920000159 gelatin Polymers 0.000 description 3
- 239000008273 gelatin Substances 0.000 description 3
- 235000019322 gelatine Nutrition 0.000 description 3
- 235000011852 gelatine desserts Nutrition 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 239000003102 growth factor Substances 0.000 description 3
- 210000003292 kidney cell Anatomy 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 229920002223 polystyrene Polymers 0.000 description 3
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 3
- 230000002441 reversible effect Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 description 3
- 229940063675 spermine Drugs 0.000 description 3
- 238000004448 titration Methods 0.000 description 3
- VVJKKWFAADXIJK-UHFFFAOYSA-N Allylamine Chemical compound NCC=C VVJKKWFAADXIJK-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 229920000742 Cotton Polymers 0.000 description 2
- ZNZYKNKBJPZETN-WELNAUFTSA-N Dialdehyde 11678 Chemical compound N1C2=CC=CC=C2C2=C1[C@H](C[C@H](/C(=C/O)C(=O)OC)[C@@H](C=C)C=O)NCC2 ZNZYKNKBJPZETN-WELNAUFTSA-N 0.000 description 2
- 102000016359 Fibronectins Human genes 0.000 description 2
- 108010067306 Fibronectins Proteins 0.000 description 2
- 102000003839 Human Proteins Human genes 0.000 description 2
- 108090000144 Human Proteins Proteins 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- KFSLWBXXFJQRDL-UHFFFAOYSA-N Peracetic acid Chemical compound CC(=O)OO KFSLWBXXFJQRDL-UHFFFAOYSA-N 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 238000005452 bending Methods 0.000 description 2
- 125000006309 butyl amino group Chemical group 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 230000001186 cumulative effect Effects 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- OSVXSBDYLRYLIG-UHFFFAOYSA-N dioxidochlorine(.) Chemical compound O=Cl=O OSVXSBDYLRYLIG-UHFFFAOYSA-N 0.000 description 2
- 239000000428 dust Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229920001002 functional polymer Polymers 0.000 description 2
- 150000004678 hydrides Chemical class 0.000 description 2
- 239000013067 intermediate product Substances 0.000 description 2
- 239000010410 layer Substances 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 235000018977 lysine Nutrition 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 230000004481 post-translational protein modification Effects 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 230000006641 stabilisation Effects 0.000 description 2
- 238000011105 stabilization Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 229960005486 vaccine Drugs 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- ZAXCZCOUDLENMH-UHFFFAOYSA-N 3,3,3-tetramine Chemical compound NCCCNCCCNCCCN ZAXCZCOUDLENMH-UHFFFAOYSA-N 0.000 description 1
- MGWGWNFMUOTEHG-UHFFFAOYSA-N 4-(3,5-dimethylphenyl)-1,3-thiazol-2-amine Chemical compound CC1=CC(C)=CC(C=2N=C(N)SC=2)=C1 MGWGWNFMUOTEHG-UHFFFAOYSA-N 0.000 description 1
- 125000004042 4-aminobutyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])N([H])[H] 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical group [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 229920000945 Amylopectin Polymers 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 0 C*(C)[C@](C=*[C+]*(*)*=C)(NC)O Chemical compound C*(C)[C@](C=*[C+]*(*)*=C)(NC)O 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 239000004155 Chlorine dioxide Substances 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 241000555268 Dendroides Species 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical group OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108700001097 Insect Genes Proteins 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- NPDTUDWGJMBVEP-UHFFFAOYSA-N N(1),N(12)-diacetylspermine Chemical compound CC(=O)NCCCNCCCCNCCCNC(C)=O NPDTUDWGJMBVEP-UHFFFAOYSA-N 0.000 description 1
- MQTAVJHICJWXBR-UHFFFAOYSA-N N(1)-acetylspermidine Chemical compound CC(=O)NCCCNCCCCN MQTAVJHICJWXBR-UHFFFAOYSA-N 0.000 description 1
- GUNURVWAJRRUAV-UHFFFAOYSA-N N(1)-acetylspermine Chemical compound CC(=O)NCCCNCCCCNCCCN GUNURVWAJRRUAV-UHFFFAOYSA-N 0.000 description 1
- GKWTZACONBQTNK-IVIZKJKWSA-N N-benzyl-7H-purin-6-amine (1R,2R,5R,8R,9S,10R,12S)-12-hydroxy-11-methyl-6-methylidene-16-oxo-15-oxapentacyclo[9.3.2.15,8.01,10.02,8]heptadecane-9-carboxylic acid (1R,2R,5R,8R,9S,10R,12S)-12-hydroxy-11-methyl-6-methylidene-16-oxo-15-oxapentacyclo[9.3.2.15,8.01,10.02,8]heptadec-13-ene-9-carboxylic acid Chemical compound C(Nc1ncnc2nc[nH]c12)c1ccccc1.CC12[C@H]3[C@H](C(O)=O)[C@@]45C[C@@H](CC[C@H]4[C@@]3(CC[C@@H]1O)OC2=O)C(=C)C5.CC12[C@H]3[C@H](C(O)=O)[C@@]45C[C@@H](CC[C@H]4[C@]3(OC1=O)C=C[C@@H]2O)C(=C)C5 GKWTZACONBQTNK-IVIZKJKWSA-N 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000005700 Putrescine Substances 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 238000004873 anchoring Methods 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000009697 arginine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000003305 autocrine Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 230000008468 bone growth Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000023402 cell communication Effects 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008614 cellular interaction Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 235000019398 chlorine dioxide Nutrition 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000004624 confocal microscopy Methods 0.000 description 1
- 239000012050 conventional carrier Substances 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 229920006037 cross link polymer Polymers 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000002242 deionisation method Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 125000004177 diethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 235000004554 glutamine Nutrition 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 229920000554 ionomer Polymers 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 125000000468 ketone group Chemical group 0.000 description 1
- 230000006651 lactation Effects 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- JCXJVPUVTGWSNB-UHFFFAOYSA-N nitrogen dioxide Inorganic materials O=[N]=O JCXJVPUVTGWSNB-UHFFFAOYSA-N 0.000 description 1
- 238000005935 nucleophilic addition reaction Methods 0.000 description 1
- 239000012434 nucleophilic reagent Substances 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000010422 painting Methods 0.000 description 1
- 230000003076 paracrine Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- JRKICGRDRMAZLK-UHFFFAOYSA-L peroxydisulfate Chemical compound [O-]S(=O)(=O)OOS([O-])(=O)=O JRKICGRDRMAZLK-UHFFFAOYSA-L 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 229920000307 polymer substrate Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- FJVZDOGVDJCCCR-UHFFFAOYSA-M potassium periodate Chemical compound [K+].[O-]I(=O)(=O)=O FJVZDOGVDJCCCR-UHFFFAOYSA-M 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- WGYKZJWCGVVSQN-UHFFFAOYSA-N propylamine Chemical group CCCN WGYKZJWCGVVSQN-UHFFFAOYSA-N 0.000 description 1
- 230000006337 proteolytic cleavage Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 229940063673 spermidine Drugs 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000000844 transformation Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 230000034512 ubiquitination Effects 0.000 description 1
- 238000010798 ubiquitination Methods 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B15/00—Preparation of other cellulose derivatives or modified cellulose, e.g. complexes
- C08B15/005—Crosslinking of cellulose derivatives
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B15/00—Preparation of other cellulose derivatives or modified cellulose, e.g. complexes
- C08B15/02—Oxycellulose; Hydrocellulose; Cellulosehydrate, e.g. microcrystalline cellulose
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B15/00—Preparation of other cellulose derivatives or modified cellulose, e.g. complexes
- C08B15/05—Derivatives containing elements other than carbon, hydrogen, oxygen, halogens or sulfur
- C08B15/06—Derivatives containing elements other than carbon, hydrogen, oxygen, halogens or sulfur containing nitrogen, e.g. carbamates
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08H—DERIVATIVES OF NATURAL MACROMOLECULAR COMPOUNDS
- C08H1/00—Macromolecular products derived from proteins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0068—General culture methods using substrates
- C12N5/0075—General culture methods using substrates using microcarriers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Other Resins Obtained By Reactions Not Involving Carbon-To-Carbon Unsaturated Bonds (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Claims (18)
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201562136241P | 2015-03-20 | 2015-03-20 | |
US62/136,241 | 2015-03-20 | ||
PCT/US2016/023248 WO2016154048A1 (en) | 2015-03-20 | 2016-03-18 | Microcarriers, matrices and scaffolds for culturing mammalian cells and methods of manufacture |
Publications (2)
Publication Number | Publication Date |
---|---|
CN107530370A true CN107530370A (zh) | 2018-01-02 |
CN107530370B CN107530370B (zh) | 2021-01-26 |
Family
ID=56978563
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201680017235.9A Active CN107530370B (zh) | 2015-03-20 | 2016-03-18 | 用于培养哺乳动物细胞的微载体、基质和支架及其制造方法 |
Country Status (6)
Country | Link |
---|---|
US (1) | US10723809B2 (zh) |
EP (1) | EP3270934B1 (zh) |
CN (1) | CN107530370B (zh) |
CA (1) | CA2980472C (zh) |
HK (1) | HK1247849A1 (zh) |
WO (1) | WO2016154048A1 (zh) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108219009A (zh) * | 2018-01-05 | 2018-06-29 | 中海石油(中国)有限公司 | 一种改性纳米纤维素及其制备方法与应用 |
CN111040983A (zh) * | 2019-12-25 | 2020-04-21 | 杭州原生生物科技有限公司 | 一种3d微载体细胞吸附培养的方法 |
CN113264868A (zh) * | 2021-05-31 | 2021-08-17 | 山东华素制药有限公司 | 一种1-苄基-3-哌啶醇的改进合成方法 |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB2544748B (en) * | 2015-11-24 | 2020-07-22 | The Electrospinning Company Ltd | Microscaffold |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4663163A (en) * | 1983-02-14 | 1987-05-05 | Hou Kenneth C | Modified polysaccharide supports |
US5766908A (en) * | 1995-03-08 | 1998-06-16 | Akzo Nobel Nv | High-flux semipermeable membrane containing immobilized affinity ligands |
Family Cites Families (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4788280A (en) * | 1987-03-19 | 1988-11-29 | National Starch And Chemical Corporation | Polysaccharide derivatives containing aldehyde groups on an aromatic ring, their preparation from the corresponding acetals and use in paper |
US5496545A (en) | 1993-08-11 | 1996-03-05 | Geltex Pharmaceuticals, Inc. | Phosphate-binding polymers for oral administration |
IL131074A0 (en) | 1999-07-23 | 2001-03-19 | Polygene Ltd | A biodegradable polycation composition for delivery of an anionic macromolecule |
DE19953589B4 (de) | 1999-11-08 | 2005-05-25 | Sca Hygiene Products Gmbh | Polysaccharid mit funktionellen Gruppen, Verfahren zu dessen Herstellung und daraus hergestellte Produkte |
DE10152407A1 (de) | 2001-10-24 | 2003-05-08 | Aesculap Ag & Co Kg | Zusammensetzung aus mindestens zwei biokompatiblen chemisch vernetzbaren Komponenten |
WO2006091894A2 (en) | 2005-02-25 | 2006-08-31 | The Regents Of The University Of California | New polymeric materials and methods |
EP2066293A2 (en) | 2006-09-29 | 2009-06-10 | Genzyme Corporation | Amide dendrimer compositions |
BRPI0720234A2 (pt) | 2006-12-14 | 2013-12-24 | Genzyme Corp | Composição farmacêutica |
US8426492B2 (en) | 2007-11-14 | 2013-04-23 | Actamax Surgical Materials, Llc | Oxidized cationic polysaccharide-based polymer tissue adhesive for medical use |
US20150239824A1 (en) | 2012-08-24 | 2015-08-27 | Universite De Strasbourg | Branched or macrocyclic polyamines and uses thereof |
EP3270933B1 (en) | 2015-03-20 | 2020-08-19 | Howard University | Polysaccharide-polyamine copolymers for removal of phosphate |
-
2016
- 2016-03-18 CN CN201680017235.9A patent/CN107530370B/zh active Active
- 2016-03-18 CA CA2980472A patent/CA2980472C/en active Active
- 2016-03-18 EP EP16769444.7A patent/EP3270934B1/en active Active
- 2016-03-18 WO PCT/US2016/023248 patent/WO2016154048A1/en active Application Filing
- 2016-03-18 US US15/560,141 patent/US10723809B2/en active Active
-
2018
- 2018-06-07 HK HK18107447.4A patent/HK1247849A1/zh unknown
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4663163A (en) * | 1983-02-14 | 1987-05-05 | Hou Kenneth C | Modified polysaccharide supports |
US5766908A (en) * | 1995-03-08 | 1998-06-16 | Akzo Nobel Nv | High-flux semipermeable membrane containing immobilized affinity ligands |
Non-Patent Citations (1)
Title |
---|
UNG-JIN KIM等: "Polyallylamine-grafted cellulose gel as high-capacity anion-exchanger", 《JOURNAL OF CHROMATOGRAPHY A》 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108219009A (zh) * | 2018-01-05 | 2018-06-29 | 中海石油(中国)有限公司 | 一种改性纳米纤维素及其制备方法与应用 |
CN111040983A (zh) * | 2019-12-25 | 2020-04-21 | 杭州原生生物科技有限公司 | 一种3d微载体细胞吸附培养的方法 |
CN113264868A (zh) * | 2021-05-31 | 2021-08-17 | 山东华素制药有限公司 | 一种1-苄基-3-哌啶醇的改进合成方法 |
CN113264868B (zh) * | 2021-05-31 | 2022-12-02 | 山东华素制药有限公司 | 一种1-苄基-3-哌啶醇的改进合成方法 |
Also Published As
Publication number | Publication date |
---|---|
EP3270934A1 (en) | 2018-01-24 |
HK1247849A1 (zh) | 2018-10-05 |
CN107530370B (zh) | 2021-01-26 |
EP3270934A4 (en) | 2018-12-12 |
US20180094080A1 (en) | 2018-04-05 |
CA2980472A1 (en) | 2016-09-29 |
CA2980472C (en) | 2023-08-29 |
US10723809B2 (en) | 2020-07-28 |
WO2016154048A1 (en) | 2016-09-29 |
EP3270934B1 (en) | 2022-12-21 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Curvello et al. | Engineering nanocellulose hydrogels for biomedical applications | |
Blanco Parte et al. | Current progress on the production, modification, and applications of bacterial cellulose | |
Courtenay et al. | Recent advances in modified cellulose for tissue culture applications | |
Markstedt et al. | Biomimetic inks based on cellulose nanofibrils and cross-linkable xylans for 3D printing | |
Grossman et al. | Biopolymer nanocomposites: processing, properties, and applications | |
Zhuang et al. | In vitro biodegradation and biocompatibility of gelatin/montmorillonite-chitosan intercalated nanocomposite | |
Dutta et al. | Chitin and chitosan: Chemistry, properties and applications | |
CN107530370A (zh) | 用于培养哺乳动物细胞的微载体、基质和支架及其制造方法 | |
Ruther et al. | Biofabrication of vessel-like structures with alginate di-aldehyde—gelatin (ADA-GEL) bioink | |
Cheng et al. | Studies on nerve cell affinity of biodegradable modified chitosan films | |
Mingyu et al. | Surface modification and characterization of chitosan film blended with poly-L-lysine | |
Unal et al. | Glioblastoma cell adhesion properties through bacterial cellulose nanocrystals in polycaprolactone/gelatin electrospun nanofibers | |
McKnight et al. | Synthesis of chitosan-alginate microcapsule membranes | |
Jämsä et al. | Versatile templates from cellulose nanofibrils for photosynthetic microbial biofuel production | |
CN110218339B (zh) | 串珠状纳米纤维素微纤维、制备方法及其在复合水凝胶制备中的应用 | |
CN104311844B (zh) | 一种基于P(MVE‑alt‑MA)光控水凝胶及其制备方法 | |
Rabbi et al. | Biocomposites of synthetic polymer modified microcrystalline jute cellulose particles and their hemolytic behavior | |
Zhang et al. | Recent advances of cellulose nanofiber-based materials in cell culture: From population to single-cell | |
CN113244453B (zh) | 可控多级交联可注射热致相变水凝胶的制备方法及应用 | |
CN108743560A (zh) | 一种磁性纤维素/β-环糊精药物可控释放水凝胶球及其制备方法 | |
CN114957730A (zh) | 一种高反应活性水凝胶微球及其制备方法和应用 | |
CN110128662A (zh) | 一种水溶性硫醇封端的peg官能化poss交联剂及其制备方法和应用 | |
Quero et al. | Production of biocompatible protein functionalized cellulose membranes by a top-down approach | |
Yokota et al. | Biofunctionality of self-assembled nanolayers composed of cellulosic polymers | |
Reiniati | Bacterial cellulose nanocrystals: production and application |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 1247849 Country of ref document: HK |
|
GR01 | Patent grant | ||
GR01 | Patent grant | ||
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20230308 Address after: 22/F, Block B, High-tech Innovation Center, Guangqiao Road, Tianliao Community, Yutang Street, Guangming District, Shenzhen City, Guangdong Province Patentee after: Shenzhen Dingsheng Biotechnology Co.,Ltd. Address before: Maryland, USA Patentee before: Yang Dazhi |
|
CI03 | Correction of invention patent | ||
CI03 | Correction of invention patent |
Correction item: Patentee Correct: Yang Dazhi False: Shenzhen Dingsheng Biotechnology Co.,Ltd. Number: 12-01 Volume: 39 |
|
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20230423 Address after: 22/F, Block B, High-tech Innovation Center, Guangqiao Road, Tianliao Community, Yutang Street, Guangming District, Shenzhen City, Guangdong Province Patentee after: Shenzhen Dingsheng Biotechnology Co.,Ltd. Address before: Maryland, USA Patentee before: Yang Dazhi |