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随着组织工程的不断发展，越来越多的学者意识到了3D培养的重要性，传统的2D培养状态并不能完全满足IPS，定向诱导多功能干细胞向组织发展，干细胞研究，再生医学，肿瘤组织药效测试，药筛和体外毒理实验的需要。3D细胞培养不同于传统在培养板或者培养皿上的2D平面贴壁培养，细胞的表达调控，外泌体分泌，细胞功能更加接近于体内的真实微环境。
本产品表面经过本公司的特殊安全无害超亲水性聚合物处里，可避免细胞附着在容器表面，进而自发性聚集于中央形成球状细胞(Spheroid Cell)。
主要特点：
1.简单的形成细胞球状体；
不需要特别步骤，只要在 96、384 孔板上播种细胞并静置培养，就能简单地获得细胞的凝集体。
2.球状体大小均等；
由于能够抑制细胞黏附在容器表面，可培养成大小均一的细胞团聚块。
3.zui适合用作分化研究；
由于能够抑制细胞黏附在容器表面，可培养成大小均一的细胞团聚块。
4.培养出的药物zui适合做抗癌药物的筛选；
通过 3D 培养，能够创造比传统的单层培养更接近生物体的环境。
实验案例：
一．使用 PrimeSurface MS-9096VZ 培养人类 ES 细胞凝集体可诱导自主形成神经视网膜组织
培养器：PrimeSurface MS-9096VZ
使用细胞： 人类 ES（KhES-1 细胞株）
接种密度：: 9,000 cells/well
使用培养基:GMEM+KSR+NEAA+2ME+ 20uM Y-27632
培养环境：5%CO2, 37℃
参考文献：・Self-Formation of Optic Cups and Storable Stratified Neural Retina from Human ESCs
NakanoT, Ando S, Takata N, Kawada M, Muguruma K, Sekiguchi K, Saito K, Yonemura S, Eiraku M, Sasai Y
Cell Stem Cell, 10 (6), 771-785 (2012)；
二．使用PrimeSurface 的抗癌药物药效的实验案例
细胞：MCF-7（人类乳腺癌细胞）
药剂：5-Fluorouracil (5-FU)
近期使用本产品发表文章：
【Retinal Research】
GAO, Lixiong, et al. Intermittent high oxygen influences the formation of neural retinal tissue from human embryonic stem cells.Scientific Reports, 2016, 6. [MS-9096V]
KUWAHARA, Atsushi, et al. Generation of a ciliary margin-like stem cell niche from self-organizing human retinal tissue. Nature communications, 2015, 6.: 1-15 [MS-9096V]
NAKANO, Tokushige, et al. Self-Formation of Optic Cups and Storable Stratified Neural Retina from Human ESCs. Cell Stem Cell, 2012, 10. 6: 771-785. [MS-9096V]
KAINI, Ramesh, et al. Xeno-free 3D retinal differentiation of human induced-pluripotent stem cells. Investigative ophthalmology visual science, 2014, 55. 13: 1369-1369. [MS-9096V]
EIRAKU, Mototsugu and SASAI, Yoshiki Mouse embryonic stem cell culture for generation of three-dimensional retinal and cortical tissues. Nature protocols, 2012, 7. 1: 69-79. [MS-9096U]
【Neuroscience Research】
FUKUSUMI, Hayato, et al. (2016). Establishment of human neural progenitor cells from human induced pluripotent stem cells with diverse tissue. Stem cells international: 1-10. [MS-9096 U, M or V], [MS-9035X, MS-9060X or MS-9090X]
RAASCH, Martin, et al. (2016). An integrative microfluidically supported in vitro model of an endothelial barrier combined with cortical spheroids simulates effects of neuroinflammation in neocortex development. Biomicrofluidics. 10: 044102. [MS-9096 U, M or V]
ISODA, Miho, et al. (2016). Robust production of human neural cells by establishing neuroepithelial-like stem cells from peripheral blood mononuclear cell-derived feeder-free iPSCs under xeno-free conditions. Neuroscience Research. [MS-9096 M]
BAMBA, Yohei, et al. (2016). In vitro characterization of neurite extension using induced pluripotent stem cells derived from lissencephaly patients with TUBA1A missense mutations. Molecular brain. 9: 70. [MS-9035X, MS-9060X or MS-9090X]
SAKAGUCHI, Hideya, et al. (2015). Generation of functional hippocampal neurons from self-organizing human embryonic stem cell-derived dorsomedial encephalic tissue. Nature communications. 6: [MS-9096 V]1-11.
TANAKA, T., et al. Generation of retinal ganglion cells with functional axons from human induced pluripotent stem cells. Sci Rep, 2015, 5. 8344. [MS-9096V]
MUGURUMA, Keiko, et al. Self-Organization of Polarized Cerebellar Tissue in 3D Culture of Human Pluripotent Stem Cells. Cell Reports, 2015, 10:537-550 [MS-9096V]
BAMBA, Y., et al. Differentiation, polarization, and migration of human induced pluripotent stem cell-derived neural progenitor cells co-cultured with a human glial cell line with radial glial-like characteristics. Biochem Biophys Res Commun, 2014, 447. 4: 683-688. [MS-9096V]
MINAMINO, Yuki, et al. Isolation and Propagation of Neural Crest Stem Cells from Mouse Embryonic Stem Cells via Cranial Neurospheres. Stem cells and development, 2014, 24.2: 172-181 [MS-9096 U, M or V]
KADOSHIMA, T., et al. Self-organization of axial polarity, inside-out layer pattern, and species-specific progenitor dynamics in human ES cell-derived neocortex. Proceedings of the National Academy of Sciences of the United States of America, 2013, 110. 50: 20284-20289. [MS-9096V]
OGAWA, Yasuhiro, et al. Impaired neural differentiation of induced pluripotent stem cells generated from a mouse model of Sandhoff disease. PLoS ONE, 2013, 8. 1: e55856. [MS-9096 U, M or V]
GOMI, Masanori, et al. Functional recovery of the murine brain ischemia model using human induced pluripotent stem cell-derived encephalic progenitors. Brain research, 2012, 1459. 52-60. [MS-9035X]
NASU, Makoto, et al. Robust formation and maintenance of continuous stratified cortical neuroepithelium by laminin-containing matrix in mouse ES cell culture. PLoS ONE, 2012, 7. 12: e53024. [MS-9096 U, M or V]
DANJO, T., et al. Subregional specification of embryonic stem cell-derived ventral encephalic tissues by timed and combinatory treatment with extrinsic signals. The Journal of neuroscience : the official journal of the Society for Neuroscience, 2011, 31. 5: 1919-1933. [MS-9096 U]
KAMIYA, Daisuke, et al. Intrinsic transition of embryonic stem-cell differentiation into neural progenitors. Nature, 2011, 470. 7335: 503-509.
KANEMURA, Yonehiro Development of cell-processing systems for human stem cells (neural stem cells, mesenchymal stem cells, and iPS cells) for regenerative medicine. The Keio journal of medicine, 2010, 59. 2: 35-45. [MS-9096 U]
【Cardiomyocytes and Heart Research】
GUO, Ge, et al. (2016). Naive pluripotent stem cells derived directly from isolated cells of the human inner cell mass. Stem Cell Reports. 6: 437-446. [MS-9096V]
NOGUCHI, Ryo, et al. (2016). Development of a three-dimensional pre-vascularized scaffold-free contractile cardiac patch for treating heart disease. The Journal of Heart and Lung Transplantation. 35: 137-145. [MS-9096 U], [MS-9035X, MS-9060X or MS-9090X]
NOGUCHI, Ryo, et al. Development of a Three-Dimensional Prevascularized Scaffold-Free Contractile Cardiac Patch for Treating Heart Disease. The Journal of Heart and Lung Transplantation, 2015, [MS-9096]
TAKASHIMA, Yasuhiro, et al. Resetting transcription factor control circuitry toward ground-state pluripotency in human. Cell, 2014, 158. 6: 1254-1269. [MS-9096V]
OTSUJI, Tomomi G, et al. Dynamic link between histone H3 acetylation and an increase in the functional characteristics of human ESC/iPSC-derived cardiomyocytes. PLoS ONE, 2012, 7. 9: e45010. [MS-9035X]
YASUDA, S., et al. A novel regulator of cardiomyogenesis in pluripotent embryonic cells. The Biochemical journal, 2011, 437. 2: 345-355. [MS-9096U]
OTSUJI, Tomomi G, et al. Progressive maturation in contracting cardiomyocytes derived from human embryonic stem cells: Qualitative effects on electrophysiological responses to drugs. Stem cell research, 2010, 4. 3: 201-213. [MS-9096U]
YAMAUCHI, Kaori, et al. Cardiomyocytes develop from anterior primitive streak cells induced by ‐catenin activation and the blockage of BMP signaling in hESCs. Genes to Cells, 2010, 15. 12: 1216-1227. [MS-9035X, MS-9060X or MS-9090X]
【Hepatocyte Research】
YANAGIDA, Ayaka, et al. Liver maturation deficiency in p57 Kip2-/-mice occurs in a hepatocytic p57 Kip2 expression-independent manner. Developmental biology, 2015, [MS-9096U]
ISHII, Takamichi (2012). Differentiation of Human Embryonic Stem Cells into Functional Hepatocyte-Like Cells (Method). Stem Cells and Cancer Stem Cells, Volume 2, Springer: 43-49. [MS-9096 U, M or V]
ISHII, Takamichi et al. (2012). Hepatic Maturation of hES Cells by Using a Murine Mesenchymal Cell Line Derived from Fetal Livers. Human Embryonic and Induced Pluripotent Stem Cells, Springer: 397-403. [MS-9096 U, M or V]
ISHII, Takamichi, et al. In vitro hepatic maturation of human embryonic stem cells by using a mesenchymal cell line derived from murine fetal livers. Cell and tissue research, 2010, 339. 3: 505-512. [MS-9096U]
【Bone and Cartliage Research】
HINO, Kyosuke, et al. (2015). Neofunction of ACVR1 in fibrodysplasia ossificans progressiva. Proceedings of the National Academy of Sciences. 112: 15438-15443. [MS-9096 U]
MURATA, Daiki, et al. A preliminary study of osteochondral regeneration using a scaffold-free three-dimensional construct of porcine adipose tissue-derived mesenchymal stem cells. Journal of orthopaedic surgery and research, 2015, 10. 1: 1-12. [MS-9096U, M or V]
FUJIMOTO, Mai, et al. Establishment of a novel model of chondrogenesis using murine embryonic stem cells carrying fibrodysplasia ossificans progressiva-associated mutant ALK2. Biochemical and Biophysical Research Communications, 2014, 455. 3: 347-352. 1. [MS-9096U, M or V]
ISHIHARA, Kohei, et al. Simultaneous regeneration of full-thickness cartilage and subchondral bone defects in vivo using a three-dimensional scaffold-free autologous construct derived from high-density bone marrow-derived mesenchymal stem cells. J Orthop Surg Res, 2014, 9. 1: 98. 1. [MS-9096U]
【Vascular Research】
KAGEYAMA, Tatsuto, et al. (2016). In situ crosslinkable gelatin-CMC hydrogels designed for rapid engineering of perfusable vasculatures. ACS Biomaterials Science Engineering. [MS-9096 U], [MS-9035X, MS-9060X or MS-9090X]
ITOH, M., et al. Scaffold-Free Tubular Tissues Created by a Bio-3D Printer Undergo Remodeling and Endothelialization when Implanted in Rat Aortae.PLoS ONE, 2015, 10. 9: e0136681. [MS-9096U]
【Dental Research】
OZEKI, Nobuaki, et al. Differentiation of Human Skeletal Muscle Stem Cells into Odontoblasts Is Dependent on Induction of 1 Integrin Expression. Journal of Biological Chemistry, 2014, 289. 20: 14380-14391. [MS-9035X, MS-9060X or MS-9090X]
YAMAMOTO, Mioko, et al. Three-dimensional spheroid culture promotes odonto/osteoblastic differentiation of dental pulp cells. Archives of oral biology, 2014, 59. 3: 310-317. [MS-9096 U, M or V]
【Somniferous Tubule Research】
YOKONISHI, T., et al. In Vitro Reconstruction of Mouse Seminiferous Tubules Supporting Germ Cell Differentiation. Biol Reprod, 2013, 89. (1):15: 1 6. [MS-9096V]
【Islet Cell Transplantation】
NAKAMURA, Kentaro, et al. (2016). Introduction to a new cell transplantation platform via recombinant peptide petaloid pieces and its application to islet transplantation with mesenchymal stem cells. Transplant International. 29: 1039-1050. [MS-9096 U]
【Bone Marrow Research】
SAYO, Kanae, et al. (2016). Fabrication of bone marrow-like tissue in vitro from dispersed-state bone marrow cells. Regenerative Therapy. 3: 32-37. [MS-9096 U]
【Generation of iPS Cell】
TAKASHIMA, Yasuhiro, et al. Resetting transcription factor control circuitry toward ground-state pluripotency in human. Cell, 2014, 158. 6: 1254-1269. [MS-9096V]
OHNISHI, Hiroe, et al. Generation of Xeroderma Pigmentosum-A Patient-Derived Induced Pluripotent Stem Cell Line for Use As Future Disease Model. Cellular Reprogramming (Formerly "Cloning and Stem Cells"), 2015, 17. 4: 268-274.
OHNISHI, Hiroe, et al. A comparative study of induced pluripotent stem cells generated from frozen, stocked bone marrow‐and adipose tissue‐derived mesenchymal stem cells. Journal of tissue engineering and regenerative medicine, 2012, 6. 4: 261-271. 1. [MS-9035X]
AOKI, T., et al. Generation of induced pluripotent stem cells from human adipose-derived stem cells without c-MYC. Tissue engineering. Part A, 2010, 16. 7: 2197-2206. [MS-9035X]
ODA, Y., et al. Induction of pluripotent stem cells from human third molar mesenchymal stromal cells. J Biol Chem, 2010, 285. 38: 29270-29278. [MS-9035X, MS-9060X or MS-9090X]
【Others】
ITO, Yoshitaka, et al. (2015). Establishment of Tsc2deficient rat embryonic stem cells. International journal of oncology. 46: 1944-1952. , [MS-9035X, MS-9060X or MS-9090X]
OGAWA, Yasuhiro, et al. (2015). Induced Pluripotent Stem Cells Generated from P0-Cre; Z/EG Transgenic Mice. PLoS ONE. 10: e0138620. [ MS-9096 U, M or V ]
IMAI, Hiroyuki, et al. Tetraploid Embryonic Stem Cells Maintain Pluripotency and Differentiation Potency into Three Germ Layers. PLoS ONE, 2015, 10. 6: e0130585. [MS-9096U]
MITSUI, Kaoru, et al. Conditionally replicating adenovirus prevents pluripotent stem cell derived teratoma by specifically eliminating undifferentiated cells. Molecular Therapy. Methods Clinical Development, 2015, 2. 15026. [MS-9096U, MS-9096M or MS-9096V]
ZHOU, Yuanshu, et al. Metabolic suppression during mesodermal differentiation of embryonic stem cells identified by single-cell comprehensive gene expression analysis. Molecular BioSystems, 2015, 11. 9: 2560-2567. [MS-9096U]
KIMURA, Kenichi, et al. The Role of CCL5 in the Ability of Adipose Tissue-Derived Mesenchymal Stem Cells to Support Repair of Ischemic Regions. Stem cells and development, 2013, 23. 5: 488-501. [MS-9096U]
SHIMOTO, Takeshi, et al. (2013). Bio Rapid Prototyping Project: Development of Spheroid Formation System for Regenerative Medicine. Information Technology Convergence, Springer: 855-862. [MS-9096U, M or V]
KOIDE, Naoshi, et al. Establishment and optimal culture conditions of microRNA-induced pluripotent stem cells generated from HEK293 cells via transfection of microRNA-302s expression vector. Nagoya journal of medical science, 2012, 74. 1-2: 157-165. [MS-9096 U, M or V]
MARKS, H., et al. The transcriptional and epigenomic foundations of ground state pluripotency. Cell, 2012, 149. 3: 590-604. [MS-9096U]
OHNISHI, Hiroe, et al. (2012). Human Mesenchymal Stem Cells and iPS Cells (Preparation Methods). Human Embryonic and Induced Pluripotent Stem Cells, Springer: 173-190. [MS-9035X, MS-9060X or MS-9090X]
SAKAI, Yusuke, et al. Embryoid body culture of mouse embryonic stem cells using microwell and micropatterned chips. Journal of bioscience and bioengineering, 2011, 111. 1: 85-91. [MS-9096U]
TAKAYAMA, Yuzo, et al. Toward the Precise Control of Cell Differentiation Processes by Using Micro and Soft Lithography. 2011, [MS-9096 U, M or V]
TAKAYAMA, Yuzo, et al. Simultaneous induction of calcium transients in embryoid bodies using microfabricated electrode substrates. Journal of bioscience and bioengineering, 2011, 112. 6: 624-629. [MS-9096U]
TANASIJEVIC, Borko and RASMUSSEN, Theodore P X chromosome inactivation and differentiation occur readily in ES cells doubly-deficient for macroH2A1 and macroH2A2. PLoS ONE, 2011, 6. 6: e21512. [MS-9096U]
KATAOKA, Ken, et al. Internalization of REIC/Dkk-3 protein by induced pluripotent stem cell-derived embryoid bodies and extra-embryonic tissues. Int J Mol Med, 2010, 26. 6: 853-859. [MS-9096U]
TAKAYAMA, Yuzo, et al. (2009). Ensemble stimulation of embryoid bodies using microfabricated ITO substrates. Engineering in Medicine and Biology Society, 2009. EMBC 2009. Annual International Conference of the IEEE, IEEE. [MS-9096U]
TAKAYAMA, Yuzo, et al. Ensemble Stimulation of Embryoid Bodies using Substrate‐Embedded Electrodes. IEEJ Transactions on Electrical and Electronic Engineering, 2009, 4. 6: 734-735. [MS-9096U]
【EST (Embryonic Stem Cell Test)】
NAGAHORI, H., et al. (2016). Prediction of in vivo developmental toxicity by combination of Hand1-Luc embryonic stem cell test and metabolic stability test with clarification of metabolically inapplicable candidates. Toxicol Lett. 259: 44-51. [MS-9096U]
YU, Ruoxing, et al. (2015). A Modified Murine Embryonic Stem Cell Test for Evaluating the Teratogenic Effects of Drugs on Early Embryogenesis. PLoS ONE. 10: e0145286. [MS-9096U]
COZ, Florian Le, et al. (2015). Hand1-Luc Embryonic Stem Cell Test (Hand1-Luc EST): A novel rapid and highly reproducible in vitro test for embryotoxicity by measuring cytotoxicity and differentiation toxicity using engineered mouse ES cells. The Journal of Toxicological Sciences. 40: 251-261.[MS-9096W]
SUZUKI, N., et al. Evaluation of novel high-throughput embryonic stem cell tests with new molecular markers for screening embryotoxic chemicals in vitro. Toxicological sciences : an official journal of the Society of Toxicology, 2011, 124. 2: 460-471. [MS-9096W]
其他研究的参考文献：
ICHIOKA, Masayuki, et al. Dienogest, a synthetic progestin, down-regulates expression of CYP19A1 and inflammatory and neuroangiogenesis factors through progesterone receptor isoforms A and B in endometriotic cells. The Journal of steroid biochemistry and molecular biology, 2015, 147. 103-110.
MORI, Taisuke, et al. Dienogest reduces HSD17 1 expression and activity in endometriosis. Journal of Endocrinology, 2015, 225. 2: 69-76.
PARSONS, Matthew W, et al. Dectin-2 Regulates the Effector Phase of House Dust Mite Elicited Pulmonary Inflammation Independently from Its Role in Sensitization. The Journal of Immunology, 2014, 192. 4: 1361-1371.
BRESLIN, Susan and O DRISCOLL, Lorraine Three-dimensional cell culture: the missing link in drug discovery. Drug Discovery Today, 2013, 18. 5: 240-249.
BARRETT, Nora A, et al. Cysteinyl leukotriene 2 receptor on dendritic cells negatively regulates ligand-dependent allergic pulmonary inflammation. The Journal of Immunology, 2012, 189. 9: 4556-4565.
MASUDA, Taisuke, et al. A microfabricated platform to form three-dimensional toroidal multicellular aggregate. Biomedical microdevices, 2012, 14. 6: 1085-1093.
SOMA, Tsutomu, et al. Hair‐inducing ability of human dermal papilla cells cultured under Wnt/ ‐catenin signalling activation. Experimental dermatology, 2012, 21. 4: 307-309.
YAMANAKA, Kaoruko, et al. Dienogest inhibits aromatase and cyclooxygenase-2 expression and prostaglandin E2 production in human endometriotic stromal cells in spheroid culture. Fertil Steril, 2012, 97. 2: 477-482.
BRENNAN, Patrick J, et al. Invariant natural killer T cells recognize lipid self antigen induced by microbial danger signals. Nature immunology, 2011, 12. 12: 1202-1211.
YOSHIIKE, Yuka and KITAOKA, Takuya Tailoring hybrid glyco-nanolayers composed of chitohexaose and cellohexaose for cell culture applications. Journal of Materials Chemistry, 2011, 21. 30: 11150-11158.
MAEKAWA, Akiko, et al. GPR17 regulates immune pulmonary inflammation induced by house dust mites. The Journal of Immunology, 2010, 185. 3: 1846-1854.
TAMADA, Atsushi, et al. Autonomous right-screw rotation of growth cone filopodia drives neurite turning. The Journal of Cell Biology, 2010, 188. 3: 429-441.
IJIMA, Hiroyuki, et al. Composition of culture medium is more important than co-culture with hepatic non-parenchymal cells in albumin production activity of primary rat hepatocytes, and the effect was enhanced by hepatocytes spheroid culture in collagen gel. BIOCHEMICAL ENGINEERING JOURNAL, 2009, 45. 3: 226-231.
ITO, Michiko and TAGUCHI, Tetsushi Enhanced insulin secretion of physically crosslinked pancreatic -cells by using a poly (ethylene glycol) derivative with oleyl groups. Acta Biomater, 2009, 5. 8: 2945-2952.
KATAOKA, M, et al. Detection of biomarker for periodontal disease using a microchip.2008,
