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INS-1 cell specific culture medium

NegotiableUpdate on 05/06

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Nature of the Manufacturer: Producers

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Overview

Related products of INS-1 cell specific culture medium: rat spleen lymphocytes, human primary outer root sheath cells, NB4 acute promyelocytic cell line, human primary peripheral blood mononuclear cells, rat colon smooth muscle cells, human primary peripheral blood mesenchymal stem cells, BJ human skin fibroblasts, human primary peripheral blood macrophages, rat colon fibroblasts, human primary coccyx tumor cells

Product Details

Product attributes:

Product Name	specification	product form
INS-1 cell specific culture medium	125mL×4	liquid

Product Details:

Carefully optimized by the technical team and tested for a long time, this product can maintainThe optimal growth state of INS-1 cells. This product already contains various ingredients required for the growth of INS-1 cells, and no additional ingredients are needed. It can be directly used for in vitro culture of INS-1 cells. This product is for further scientific research use only and should not be used for diagnosis, treatment, clinical, household, or other purposes.

Product Description

product form: Liquid

product concentration:1×

Product Specifications:125mL×4

Composition of culture medium:RPMI-1640＋10% FBS＋50μmol/L β-mercaptoethanol＋1% P/S

Bacterial detection: Negative

Fungal detection: Negative

Mycoplasma testing: Negative

Cell Growth ExperimentGood cell growth and normal morphology

Endotoxin content(EU/mL):≤3

Storage conditions: 2-8 ℃, stored away from light

transport conditionsIce bag refrigerated transportation

Validity period: 3 months

Experimental report:

1、 Separation and cultivation:

2、 Immunofluorescence identification:

1. Under sterile conditions, extract atrial tissue from 1-3 day old SD rats, wash the tissue block twice with PBS, and finally reduce it to a size of approximately 1mm3;
2. Add 4 mL of enzyme digestion solution (0.1% and 0.1% type I collagenase) to the tissue block, suspend for 10 seconds, digest at 37 ℃ for 10 minutes, then use a dropper to prepare a single-cell suspension, naturally precipitate and collect the supernatant, terminate digestion with 10% FBS medium, and place at 4 ℃;
3. Add 3-4mL of enzyme digestion solution to the remaining tissue, suspend for 10 seconds, digest at 37 ℃ for 10 minutes, collect the supernatant according to the above method, terminate digestion, and place at 4 ℃. Repeat this step 2-3 times until the tissue is digested;
4. Filter the cell digestion solution through a 200 mesh stainless steel sieve, centrifuge at 1200r/min for 10 minutes, discard the supernatant, and suspend the precipitated cells in DMEM/F12 medium containing 10% FBS. Inoculate the cells into a 25cm2 culture bottle and incubate in a 37 ℃, 5% CO2 incubator;
5. After 1 hour of differential adhesion, aspirate the culture medium and inoculate it into a 6-well plate as needed for further cultivation;

1. When the atrial myocytes grow to 80% fusion, discard the culture medium and wash the cells twice with warm PBS for 10 minutes each time. Then fix the cells with 4% paraformaldehyde at room temperature for 15 minutes;
2. Wash the cells twice with PBS for 10 minutes each time, and then permeate the membrane with 0.1% Triton X-100 at 4 ℃ for 15 minutes;
3. Wash the cells twice with PBS for 10 minutes each time, and then block the cells with 4% BSA at room temperature for 30 minutes;
4. Dilute the alpha actin primary antibody in a ratio of 1:100, and then incubate the cells overnight at 4 ℃ in a refrigerator;
5. Wash the cells with PBS three times, each time for 10 minutes. Dilute the secondary antibody against alpha actin in a ratio of 1:150 and place it at 37 ℃ for 1 hour;
6. Wash with PBS three times, each time for 10 minutes, and finally observe the image under an inverted fluorescence microscope and take photos.

Operating procedures:
1、 Preparation of culture medium and culture cryopreservation conditions:
1) Prepare RPMI-1640 medium, 90%; High quality fetal bovine serum, 10%.
2) Cultivation conditions: Gas phase: air, 95%; Carbon dioxide, 5%. Temperature: 37 ℃, humidity in the incubator is 70% -80%.
3) Cryogenic solution: 90% serum, 10% DMSO, ready to use and prepared, stored in liquid nitrogen.
2、 Cell processing:
1) Resuscitate cells: Quickly place a cryovial containing 1mL of cell suspension into a 37 ℃ water bath (with the water level lower than the lid of the cryovial) and shake to thaw. Transfer the cryovial into a 15ml centrifuge tube containing 4mL of culture medium prepared in advance and mix well. Centrifuge at 1000RPM for 4 minutes, discard the supernatant, add 1mL of culture medium and blow evenly. Then transfer all cell suspensions into a culture bottle containing 5ml of culture medium and culture overnight. The next day, change the solution and check the cell density.
2) Cell passage: If the cell density reaches 80% -90%, passage culture can be carried out.
For suspended cells, the following methods can be used for passaging:
Method 1: Collect cells,1000RPM， Centrifuge at room temperature for 5 minutes, discard the supernatant, add 1-2mL of culture medium and blow evenly. Divide the cell suspension into new dishes or bottles containing 8ml of culture medium in a ratio of 1:2 to 1:5.
The company's products are only used for scientific research

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