hPSC Cardiac Organoid Differentiation Kit
hPSC Cardiac Organoid Differentiation Kit
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| Usage | Use low adsorption 96 Well plate, inducible 2 board 192 cardiac organoids, success rate 70-80% 。 I. Experimental Instruments and Materials Instruments: Biosafety cabinet, cell incubator, horizontal centrifuge, inverted microscope, cryogenic refrigerator Materials: Cell culture plates (specifications are: 96 Hole), centrifuge tube (specifications are 15mL 、 50mL ), pipette (specifications are 10uL 、 100 uL 、 1000 uL ), sterile tip (specifications are 10 uL 、 200 uL and 1000 uL ）、 Pipette (size is 10mL 、 50mL ） II. Experimental Methods 1 、 Ⅰ Stage ( D0-2 ) (Press 1 one 96 Well plate, per well 100uL (Calculate) （ 1 ） hPSC One hole in the six-well plate coated with matrix glue grows to 70-80% Convergence degree. （ 2 Remove the culture medium. （ 3 Add to the hole 1mLPBS (No calcium or magnesium ions) Wash and absorb. （ 4 Add to the hole 1mLAccutase ， 37℃ digestion 3min 。 （ 5 After digestion, the clone can be seen to be white and shiny under a microscope. Gently tap the sides of the well plate a few times to detach the cells from the bottom of the plate, then add... 1mLhESC/iPSC Complete medium was terminated for digestion and used P-1000 The suction head will 2mL Cell monosuspension was collected 15mL In a centrifuge tube, at room temperature 300g centrifuge 5min 。 （ 6 After centrifugation is complete, remove the supernatant to remain. 20ul Gently flick the bottom of the pipe 3-4 Next, will 1mL preheated hESC/iPSC Complete culture medium (including 10μMY-27632 Resuspend the cells and gently pipette up and down to ensure uniformity of the single-cell solution. （ 7 Cells were counted using an automated cell counter, and trypan blue was used to exclude dead cells. , per hole 5000 The desired cells were collected into centrifuge tubes based on the proportion of cells, and the basal culture medium was added. 1 With supplements A Mix thoroughly, and combine the 1mL cell suspension addition 9mL contains supplements A basic culture medium 1 In the middle, blow the cells evenly, then press each well. 100uL The volume of the cell suspension was inoculated into 96 In the orifice plate, 1000rpm centrifuge 5min Then transfer the well plate to 37℃ 、 5%CO2 Incubation in cell culture box 3 heaven. 2 、 Ⅱ Stage ( D3-4 ） No 3 Days, the basic culture medium 2 With supplements B Mix thoroughly and configure to Ⅱ Stage culture medium: Aspirate the medium from the wells, per well. 100uL The volume will Ⅱ Stage culture medium is added to the wells, and the culture plate is placed in... 37℃5%CO2 Incubator culture, continuous culture 2 heaven. 3 、 Ⅲ Stage ( D5-6 ） No 5 Days, the basic culture medium 3 With supplements C Mix thoroughly and configure to Ⅲ Stage culture medium: Aspirate the medium from the wells, per well. 100uL The volume will Ⅲ Stage culture medium is added to the wells, and the culture plate is placed in... 37℃5%CO2 Incubator culture, continuous culture 2 heaven. 4 、 Ⅳ Stage ( D7-21 ） （ 1 )th 7 During the day, the culture medium was aspirated from the wells, and each well was aspirated. 100uL The volume of the basic culture medium 4 Add to the well and place the culture plate in 37℃5%CO2 Cultivate in an incubator, beginning long-term cultivation, each 2 Change the culture medium once a day. （ 2 Cardiac organoids will undergo differentiation. 10-13 The sky began to dance, and after further cultivation... 19-21 Daily sampling and identification.
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| Description | This product is hPSC Cardiac organoid kit for inducing differentiation, human pluripotent stem cells hPSC Cardiac organoids can be obtained by inducing differentiation through the kit. The organoids are composed of cardiomyocytes, epicardial cells and other cells. The mature organoids have a chamber structure and early beating function. This kit requires the operator to have hPSC Cultivation experience and a certain understanding of organoids. Product Composition:
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| Storage Temp. | See product composition for details. |