{"product_id":"hpsc-derived-whole-brain-organoid-kit-abs90473","title":"hPSC 诱导分化全脑类器官试剂盒","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 196 For individual brain organoids, it is recommended that each induction... 1\/2 The six-well plate initiates the differentiation process, forming at least 98 uniform size EB The ball undergoes a whole-brain induction process. \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 ）、 10uL\/100uL 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 embryo-like body (EBs) Formation (th 0-2 (sky) \u003c\/strong\u003e\u003cbr\u003e Preparation of Embryoid Formation Medium \u003cbr\u003e Embryoid formation medium ( 12mL\/3 hole) : Recovering to room temperature before use, 12mL Basic medium 1+1x24uL supplements A Ready to use. at room temperature ( 15-25°C Thaw the supplement and mix thoroughly. Note: If not used immediately, at most... 2-8°C store 2 Weeks, do not exceed the shelf life of the basic medium and supplements, use immediately after thawing, do not freeze and thaw repeatedly. If the supplement is not mixed, put the supplement in -20°C Save. Do not exceed the shelf life of the supplement. Use immediately after thawing. Do not freeze or thaw repeatedly. \u003cbr\u003e①Day0-2\u003cbr\u003e1 Before use, restore the culture dish, medium, and reagents to room temperature. 15-25°C ）。 \u003cbr\u003e2 ,when iPSC\/hESC Grow to 70-80% Confluence degree, microscopic observation of cells in good condition. \u003cbr\u003e3 For 3 hole \/6 orifice plate 80% density iPSC\/hESC Cells, prepare embryo-like formation medium ( 12mL\/3 hole). prepare 3mLAccutase 、 50mLDPBS Sterile calcium-magnesium-free, and 10mLDMEM\/F12 Preheat to room temperature. \u003cbr\u003e4 Use room temperature without calcium or magnesium DPBS Wash all three holes of the six-hole plate once and discard. DPBS ， s Add per well 1mL of Accutase37 degree of digestion 3min 。 \u003cbr\u003e5 When you see the clone turn white and shiny, gently tap the side of the plate (twice on each side) to take a picture of the cells, and then add them to the biosafety cabinet. 1mLDMEM\/F12 Terminate digestion, blow until cells are completely detached and form a single cell type, and trypan blue counts are performed. Cell viability is higher than... 90% The next step of induction can then be carried out. \u003cbr\u003e6 、 160g\/5min Collected by centrifugation 6mL Cell suspension, discard the supernatant and add 12mL embryoid formation medium , Cell viability and cell count were tested again, and finally each mL The number of cells in 50000-80000 The best between them. \u003cbr\u003e7 Prepare an ultra-low adsorption 96 hole U The base plate will 12mL Cell suspension (containing supplement) A Add it to the sampling tank and use a row gun. \/ Add to each well of the pipette 100uL The cell suspension. After sample addition, seal the petri dish with a sealing film and use a horizontal plate centrifuge. 1200rpm\/3min Centrifugation. \u003cbr\u003e8 Tear off the sealing membrane and place it in a cell culture incubator for overnight culture. Add it to each well the next day. 100uL Basic medium 1 。 \u003cbr\u003e9 、 48 After an hour, the suspended cells can aggregate into spheres with smooth and rounded edges. EBs In optimal condition. Subsequently, the next experimental operation was performed. \u003cbr\u003e Note: \u003cbr\u003ePSC 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\u003e\u003cdiv\u003e\n\u003cstrong\u003e\u003cbr\u003e Neuroectodermal differentiation (th 2-6 (sky) \u003cbr\u003e\u003c\/strong\u003e Return the culture dish, medium and reagents to room temperature before use ( 15-25°C ）。 \u003cbr\u003e①Day2-6\u003cbr\u003e1 , will induce well EBs Transfer to ultra-low adsorption 6 In the hole plate, add to each hole 10 one EBs Use later DPBS Wash once to remove any medium residue from the previous stage, then discard. DPBS Then add to each well 2ml basic culture medium 2 , every day 2 The fluid is changed every other day to induce co- 4 heaven. \u003cbr\u003e2 Neuroectodermal expansion occurs 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\u003e\u003cbr\u003e\u003cstrong\u003e Nerve budding (No. 6-25 (sky) \u003c\/strong\u003e\u003cbr\u003e Return the culture dish, medium and reagents to room temperature before use ( 15-25°C ）。 \u003cbr\u003e①Day6-25\u003cbr\u003e1 ,when EBs When the diameter meets the relevant standards 400-600um Can be embedded with matrix rubber, 4 Low growth factor matrix glue for freezing and dispensing 2ml 。 \u003cbr\u003e2 Prepare a sheet of equal length and width. 10cm The sealing film, in 200ul Press a dent into the gun barrel and place it. 10cm The dish was soaked in alcohol. 30 Seconds later, place in a safe cabinet for ultraviolet drying. 30min The above can be used. \u003cbr\u003e3 Use a wide mouth 10-100uL pipetting gun tip ( measuring range 50uL) Absorption of the end of neuroectodermal differentiation EB On the concave points pressed, it is recommended to operate once. 10 One is preferable. Afterwards, a pipette was used to carefully absorb any remaining culture medium from the pits, and each pit and... EBs join 30uL Careful embedding of low growth factor matrix gel EBs , if EBs At the edge of the colloid, use 10uL The little spearhead was gently prodded EBs The surrounding colloid places it in the center. ( Be careful not to form bubbles. ) 。 \u003cbr\u003e4 The buried EBs Cover it and place it in 37 degree gel 15min Then, use sterile tweezers to lift the sealing membrane. 1mL basic culture medium 3 Each row EBs The colloid is blown against the bottom of the concave point until the adsorption is ultra-low. 6 In the orifice plate, each orifice can hold a maximum of 10 For organoids, the volume of culture medium per well should not exceed 3mL 。 \u003cbr\u003e5 If too many are made at once EBs You can also directly pre-cool it. 30min The above 12mL basic culture medium 3 supplement 10% Low growth factor matrix gel, (i.e. 1 、 2mL After the colloid is mixed evenly in a vortex. Aspiration of the basal medium in the original well plate 2 Replenish each well 3mL Basic medium 3+ Low growth factor matrix gel, induced 48 After an hour, the medium was aspirated and used... DPBS Wash twice and add again. 3mL Base medium without matrix gel per well 3 Conduct the first 6 Step-by-step operation. \u003cbr\u003e6 Place the ultra-low six-well plate on a horizontal shaker. (80rpm\/s) inside 37 degree cultivation 18 The expansion of the neuroepithelium was completed every two days, and the fluid was changed every two days. EBs Generally in the neuroepithelial dilation stage 4 Early appearance during the day VZ 、 SVZ Such a structure, 12 The sprouts gradually converge and turn black. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e Neurological and Brain Maturation (Part 1) 25 (Power Queen) \u003c\/strong\u003e\u003cbr\u003e Return the culture dish, medium and reagents to room temperature before use ( 15-25°C ）。 \u003cbr\u003e①Day25+\u003cbr\u003e1 ,when EBs The center is dense, the outer layer forms a loose epithelial structure with a diameter greater than 1mm This indicates that the nerves have expanded to the mature stage and the basal culture medium has been completely abandoned. 3 Add to each well 3mL basic culture medium 4 Continuous training until 38 Pop Queen ( Change the fluid every two days \/80rpm\/s) ， EBs The diameter is greater than 2mm At this time, nutrients may not penetrate the interior, requiring the use of... 1mL The syringe will EBs Carefully remove the surrounding colloid and add it to each well. 3mL Basic medium 5( Change the fluid daily ) And increase the shaker speed to 120rpm\/s Improving nutrient circulation allows brain organoids to be cultured for a long time. 100 heaven. \u003cstrong\u003e\u003cbr\u003e\u003c\/strong\u003e\n\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 Whole brain organoid kit for inducing differentiation, human pluripotent stem cells hPSC Whole-brain organoids can be obtained by inducing differentiation through the kit. The kit completes the classic brain organoid differentiation process through four components. : Neuroectodermal differentiation, neuroepithelial sprouting and neural tube formation, nerve expansion, brain maturation. The induction process requires implantation EB Ball on low growth factor matrix gel, inducing 38 Whole-brain organoid expression for more than 10 days TUJ1 、 SOX2 、 Nestin 、 NeuN Isobasic neural characterization, FOGX1 、 CTIP2  Isoforebrain and cortical characterization , After entering long-term cultivation ≥80 Small amounts can be expressed daily GFAP 、 TH Isofunctional neurons and glial cells Maker 。 \u003cbr\u003e This kit requires the operator to have hPSC Cultivation experience and a certain understanding of organoids. \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: 161px; 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: 44px;\" rowspan=\"2\"\u003e embryo-like body formation stage \u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e Basic medium 1\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e200 mL\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: 22px;\"\u003e supplements A 500X\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 22px;\"\u003e400 uL\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=\"height: 25px; width: 18.892%;\"\u003e Neuroectodermal differentiation stage \u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 25px;\"\u003e Basic medium 2\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 25px;\"\u003e100mL\u003c\/td\u003e\n\u003ctd style=\"width: 18.892%; height: 25px;\"\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 nerve budding differentiation 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 stage of neural maturation and differentiation \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;\"\u003e200mL\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 neural maturation maintenance 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: 286px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; height: 22px; text-align: center;\"\u003e Product Name \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003e Source \u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; 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: 20.6132%; height: 22px; text-align: center;\"\u003ehESC\/iPSC cell culture kit \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; height: 22px; text-align: center;\"\u003eabs90487\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; height: 22px; text-align: center;\"\u003e stem cell matrix glue \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; height: 22px; text-align: center;\"\u003eabs9496\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; height: 22px; text-align: center;\"\u003e DMEM\/F-12\u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; height: 22px; text-align: center;\"\u003eabs9560\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; height: 22px; text-align: center;\"\u003e low growth factor matrix glue \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003e-\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; height: 22px; text-align: center;\"\u003e- \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; height: 22px; text-align: center;\"\u003e Dulbecco's phosphate buffer (DPBS) ， \u003cbr\u003e Does not contain calcium, magnesium, phenol red \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; height: 22px; text-align: center;\"\u003eabs970\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; height: 22px; text-align: center;\"\u003e Alcohol \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003e-\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; height: 22px; text-align: center;\"\u003e-\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; height: 22px; text-align: center;\"\u003e6 Aperture plate ( TC dispose \/ (Non-low adsorption) \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; height: 22px; text-align: center;\"\u003eabs7033\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; height: 22px; text-align: center;\"\u003e6 hole ultra-low adsorption plate \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; height: 22px; text-align: center;\"\u003eabs7056\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; height: 22px; text-align: center;\"\u003e ultra-low adsorption 96 hole U bottom plate \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; height: 22px; text-align: center;\"\u003eabs7060\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; text-align: center; height: 22px;\"\u003e Accutase\u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; text-align: center; height: 22px;\"\u003eabs47014935\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; text-align: center; height: 22px;\"\u003e10uL\/100uL wide-mouth pipette tip \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; text-align: center; height: 22px;\"\u003eabs7075\/abs7079\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 20.6132%; text-align: center; height: 22px;\"\u003e Taipan Blue \u003c\/td\u003e\n\u003ctd style=\"width: 23.9324%; text-align: center;\"\u003eAbsin\u003c\/td\u003e\n\u003ctd style=\"width: 19.7151%; text-align: center; height: 22px;\"\u003eabs50036\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\u003ePlease refer to the 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":43224458887243,"sku":"abs90473-1kit","price":4603.0,"currency_code":"USD","in_stock":true}],"url":"https:\/\/www.antbioinc.com\/products\/hpsc-derived-whole-brain-organoid-kit-abs90473","provider":"AntBio","version":"1.0","type":"link"}