{"product_id":"hpsc-derived-cortical-brain-organoid-kit-abs90474","title":"hPSC 诱导分化皮质脑类器官试剂盒","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e This kit specification can produce more than 200 For individual brain organoids, it is recommended that each induction... 1 The six-well plate initiates the differentiation process, forming at least 60-100 A uniformly sized epineural balloon (ideal formation rate) 60% ）。 \u003cbr\u003e\u003cstrong\u003e I. Experimental Instruments and Materials \u003cbr\u003e\u003c\/strong\u003e Instruments: Biosafety cabinet, cell incubator, horizontal centrifuge, inverted microscope, cryogenic refrigerator \u003cbr\u003e Material: Centrifuge tube (specifications are 15mL and 50mL ), pipette (specifications are 10uL 、 100uL 、 1000uL and multi-channel 100uL ), sterile tip (specifications are 10uL 、 200uL and 1000uL ), pipette (specifications are 10mL 、 50mL ）、 1mL\/200uL a wide-mouth pipette tip, 6 Aperture plate ( TC dispose \/ non-low adsorption), 6 Pore ultra-low adsorption plate, ultra-low adsorption 96 hole U bottom plate \u003cbr\u003e\u003cstrong\u003e II. Experimental Methods \u003cbr\u003e\u003c\/strong\u003ehESC\/iPSC cell preparation \u003cbr\u003e In 6 Aperture plate ( TC dispose \/ (Non-low adsorption) hESC\/iPSC The cell culture kit will hESC\/iPSC Cell culture to confluence 70-80% 。 \u003cbr\u003e\u003cstrong\u003e Neuroepithelial differentiation (two modes of differentiation) \u003c\/strong\u003e\u003cbr\u003e① neuroepithelial stem cells NESC two-dimensional induction \u003cbr\u003e1 Before use, restore the culture dish, medium, and reagents to room temperature. 15-25℃ ）。 \u003cbr\u003e2 ,when iPSC\/hESC Two days after passaging, neuroepithelial induction was performed directly, and the cell morphology was observed to form small island-like clones, with a cell clone density lower than 30% It can be induced without single-cell stress growth and stressed globular clones. Discard hESC\/iPSC Use of cell culture kit DMEM\/F12 The cells were washed twice to remove floating cells and then added to each well. 2mL Basic medium 1 , each 2 The solution is changed daily until the cell clone density reaches... 80%,10x Under a microscope, the clone edges were rounded and smooth, the cell nuclei were smaller and more numerous, and the induction time meter was observed. 3-4 heaven. \u003cbr\u003e Note: PSC In case of cloning problems, self-transformation, or floating, resuscitation and amplification of one plant should be initiated again. It should not be used directly for induction. \u003cbr\u003e② .from EB Direct neuroepithelial induction of the sphere (this method is recommended for differentiation by beginners) )\u003cbr\u003e1 Before use, restore the culture dish, medium, and reagents to room temperature. 15-25℃ ）。 \u003cbr\u003e2 ,will iPSC\/hESC Cell culture to 80% The procedure can be performed as long as the clone density, clear and rounded clone boundaries, and normal nuclear-plasmid ratio are obtained. EB induce, DPBS After washing the cells twice, each well was washed with 500uL of Accutase or hESC\/iPSC Passage working solution (enzyme-free), digested 6-10 After 15 minutes, it was observed that the clone became rounded, shrunk, and brightened, and when shaken, a small number of cells detached from the matrix, and digestion was terminated. \u003cbr\u003e3 Absorb digestive juices, if used Accutase If digestion causes a large number of cells to float, they will not be absorbed; an equal amount will be used. DMEM\/F12 Termination of digestion. And hESC\/iPSC After passage working solution (without enzyme) is digested, it is directly added to each well after aspiration. 500uL Basic medium 2 After resuspending the cells, they were 15mL or 50mL In a centrifuge tube (flexibly selected according to the type of digestive fluid). \u003cbr\u003e4 、 Accutase Digested cells 120g centrifuge 5 After discarding the supernatant for minutes, add the number of digested wells. *500uL Add the volume of basal medium 2 Resuspend and replenish 1X supplements A , and hESC\/iPSC Cells digested with passage working solution (enzyme-free) were directly added to the original resuspension system (number of digestion wells). *500uL The volume of basal culture medium 2 Later supplement 1x supplements A 。 \u003cbr\u003e5 The cell suspension +1x supplements A Add to the sampling tank or 10cm Cell culture dishes, using a row gun per well 100uL Add ultra-low adsorption 96 hole u In the bottom petri dish, after the sealing film is wrapped, a horizontal plate removal machine is used. 1500rpm centrifuge 5 minutes, 37 The cells were cultured overnight in a cell incubator, and added to each well the next day. 100uL Basic medium 2( No supplements A) ， EB Total formation time approx. 48 Hours. \u003cbr\u003e6 On the third day, the culture medium was completely aspirated and added to each well. 200uL Basic medium 1 Cultivate for three days, then absorb on the second day 100uL rehydration 100uL Removal of culture metabolic waste. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e Neurosphere formation (directly from EB Methods for inducing neuroepithelium are not referenced. 2-4 Look directly 5)\u003c\/strong\u003e\u003cbr\u003e1 Before use, restore the culture dish, medium, and reagents to room temperature. 15-25℃ ）。 \u003cbr\u003e2 ,will 6mLAccutase After warming to room temperature, place in a biosafety cabinet and use in each well. 1mL of DPBS Induced neuroepithelial stem cells were washed twice and added to each well. 800uL-1000uL of Accutase Place in an incubator for digestion. 5-8 Minutes, every 4 The cell state is observed once every minute. The cell clone edges are curled, the cell spaces become bright, and a small number of cells float in the digestive fluid to terminate digestion. \u003cbr\u003e3 Digestive fluid is drawn from the side wall and added to each well. 1mL basic culture medium 3 After resuspending the cells, they were concentrated 12mL basic culture medium 3+12uL of 1x supplements A If the cells are overdigested and most of them float completely, then use each well. 1mL of DMEM\/F12 After termination of digestion 120g\/3min Cells were collected by centrifugation and then concentrated in 12mL basic culture medium 3+12uL1x supplements A Attention : Before dilution, trypan blue staining detected cell activity higher than 80% You can then proceed to the next step. Neuroectodermal expansion EB The edges become transparent, and the diameter is \u0026gt; 400um The interior is dense and cavity-free, and the overall shape is similar to that of a primary mouse neurosphere. \u003cbr\u003e4 After centrifugation, cells should be gathered on one side of the well and placed in an incubator for culture. 24h Then add more to each well. 100uL basic culture medium 3 No supplements A Neurosphere formation induction time meter 2 heaven. \u003cbr\u003e5 For direct formation EB The post-induction neuroepithelial system is completed. 3 The basal culture medium was completely absorbed after induction. 1 Add per well 250uL Basic medium 3 induced 2 heaven. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e Nerve rosette expansion ( (Both schemes are consistent) \u003c\/strong\u003e\u003cbr\u003e1 Before use, restore the culture dish, medium, and reagents to room temperature. 15-25℃ ）。 \u003cbr\u003e2 When the neurosphere diameter meets the relevant standards ( 200-600μm ), use a wide mouth 200uL The pipette tip aspirates the neurosphere for ultra-low adsorption. 6 In the hole plate, each hole can accommodate a maximum of 10 A neural sphere, so at least prepare 2 The six plates initiated the expansion of the nerve lotus base. \u003cbr\u003e3 Neurosphere entry 6 After the hole plate is in the middle, gently add each hole. 2mL Basic medium 4 Then, place the cross. \/ rotary shaker 80rpm\/s upper cultivation 18 Day, every 1-2 Change the fluid every day \/ Liquid change per well 2mL 。 \u003cbr\u003e\u003cdiv\u003e\n\u003cstrong\u003e\u003cbr\u003e cortical brain maturation \u003cbr\u003e\u003c\/strong\u003e1 Before use, restore the culture dish, medium, and reagents to room temperature. 15-25℃ ）。 \u003cbr\u003e2 When black spotty structures appear in the center and outer sides of the neurosphere, it indicates that the neural rosette has expanded to a suitable stage. Completely discard the culture medium and add it to each well. 2mL basic culture medium 5 Place the cross \/ rotary shaker 80rpm\/s upper cultivation 18-26 sky \/ Every 2-3 A single liquid change can be used for identification and downstream experiments. Structurally mature brain organoids IF Staining visible TUJ1\/SOX2 Labeled cell formation VZ and SVZ It has a similar structure and high expression of glutaminergic-related genes, using basic culture medium. 5 Long-term shaker culture can cultivate to 120 No apoptosis occurred in the central cells. \u003cbr\u003e3 When the cortical brain is cultured to a diameter greater than 1mm In the above cases, it is necessary to cut it to reduce internal cell apoptosis. Use sharp, blunt edges 2 The surgical blade will put a 1mm The above cortical organoids are cut into 2-4 block, DPBS After washing the organoids once, use a wide-mouthed 1000uL The pipette tip was transferred to 15mL Natural sedimentation in the centrifuge tube (approximately 5-8min ), absorb DPBS After supernatant, add basal medium 4+1x supplements A Transfer into ultra-low adhesion six-well plate 80rpm\/s cultivate 4 Change the fluid every day. 4 After a few days, add and change to basic culture medium. 5120rpm\/s Continuing to maintain culture can prolong culture time and organoid activity to 180 heaven. \u003c\/div\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e This product is hPSC Cortical brain organoid kit for inducing differentiation, human pluripotent stem cells hPSC Cortical brain organoids can be obtained by inducing differentiation through the kit, and the kit is a standardized culture system designed specifically for simulating human brain cortex development and function research through forebrain ventralization signal pathway regulation technology. The system is activated through timing. Wnt\/β-catenin and gradient suppression BMP\/Smad Signals drive human pluripotent stem cells ( PSC Directed differentiation into high-purity glutamatergic neurons ( VGLUT1\/2 positive rate \u0026gt;85% ), and simultaneously induce radial glial cells ( Pax6+\/BLBP+ This forms a biomimetic ventricular structure. Available in 60 Intraday stable formation diameter 1.5-2.0mm Lamellar cortical organoids that characterize the expression of deep cortical markers TBR1 (th Ⅴ-Ⅵ layer) and surface layer SATB2+ Neurons (th Ⅱ-Ⅳ (layer), and accompanied by MBP+\/MOG+ Oligodendrocyte-mediated dense myelination ( LFB staining positive rate \u0026gt;70% ）。 This system uses a three-dimensional rotating bioreactor or ultra-low adhesion well plate ( 3DRBC The oxygen gradient is maintained at 8-12% Achieve high-throughput culture (single batch) ≥96 organoids). This model is suitable for neurodegenerative diseases (such as Alzheimer's disease). tau Protein pathology modeling), schizophrenia NMDA Research on receptor function resolution and synaptic plasticity in autism spectrum disorders, while compatible with single-cell sequencing and calcium imaging dynamic tracking technologies. \u003cbr\u003e This kit requires the operator to have hPSC Cultivation experience and a certain understanding of organoids. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e\u003cstrong\u003e Product Composition: \u003cbr\u003e1 Kit contents \u003cbr\u003e\u003c\/strong\u003e\u003c\/strong\u003e\u003ctable style=\"border-collapse: collapse; width: 95%; height: 156px; margin-left: auto; margin-right: auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e Research Phase \u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e Product Name \u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e Product Specifications \u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e store \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 18.892%; height: 39px;\" rowspan=\"3\"\u003e Neuroepithelial differentiation stage \u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 17px;\"\u003e Basic medium 1\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 17px;\"\u003e200mL\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 17px;\"\u003e-20℃ ， 24 Month \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e Basic medium 2\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e100mL\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e-20℃ ， 6 Month \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 18.892%; height: 25px;\"\u003e supplements A500X\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 25px;\"\u003e400uL\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 25px;\"\u003e-20℃ ， 6 Month \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e neurosphere formation stage \u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e Basic medium 3\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e200mL\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e-20℃ ， 24 Month \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"height: 26px; width: 18.892%;\"\u003e Nerve rosette expansion stage \u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 26px;\"\u003e Basic medium 4\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 26px;\"\u003e400mL\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 26px;\"\u003e-20℃ ， 24 Month \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e Cortical brain maturation stage \u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e Basic medium 5\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e200mL\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e-20℃ ， 24 Month \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003e\u003cstrong\u003e\u003cstrong\u003e2 Other related reagents and consumables \u003cbr\u003e\u003c\/strong\u003e\u003c\/strong\u003e\u003ctable style=\"border-collapse: collapse; width: 100%; height: 264px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003e Product Name \u003c\/td\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003e Source \u003c\/td\u003e\n\u003ctd style=\"width: 32.1308%; height: 22px; text-align: center;\"\u003e Product number \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003ehESC\/iPSC cell culture kit \u003c\/td\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 32.1308%; height: 22px; text-align: center;\"\u003eabs90487\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003e stem cell matrix glue \u003c\/td\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 32.1308%; height: 22px; text-align: center;\"\u003eabs9496\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003eDMEM\/F-12\u003c\/td\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 32.1308%; height: 22px; text-align: center;\"\u003eabs9560\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003eDulbecco's phosphate buffer (DPBS) ， \u003cbr\u003e Does not contain calcium, magnesium, phenol red \u003c\/td\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 32.1308%; height: 22px; text-align: center;\"\u003eabs970\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003ehESC\/iPSC Passage working solution (enzyme-free) \u003c\/td\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 32.1308%; height: 22px; text-align: center;\"\u003eabs90493\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003e6 Aperture plate ( TC dispose \/ (Non-low adsorption) \u003c\/td\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; 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height: 22px; text-align: center;\"\u003eAccutase\u003c\/td\u003e\n\u003ctd style=\"width: 32.1299%; height: 22px; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 32.1308%; height: 22px; text-align: center;\"\u003eabs47014935\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.1299%; text-align: center; height: 22px;\"\u003e Taipan Blue \u003c\/td\u003e\n\u003ctd style=\"width: 32.1299%; text-align: center; height: 22px;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 32.1308%; text-align: center; height: 22px;\"\u003eabs50036\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.1299%; text-align: center; height: 22px;\"\u003e200uL\/1mL wide-mouth pipette tip \u003c\/td\u003e\n\u003ctd style=\"width: 32.1299%; text-align: center; height: 22px;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 32.1308%; text-align: center; height: 22px;\"\u003eabs7075\/abs7079\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eSee product composition for details.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"1kit","offer_id":43224459083851,"sku":"abs90474-1kit","price":4603.0,"currency_code":"USD","in_stock":true}],"url":"https:\/\/www.antbioinc.com\/products\/hpsc-derived-cortical-brain-organoid-kit-abs90474","provider":"AntBio","version":"1.0","type":"link"}