Hi Heena,

Beneath please find a protocol for cultures using 6 well plate you required. Upon receipt of cultures, the following describes

procedures for plating a single cell suspensions in 6 well plates, for

Tumorsphere formation.

Other Equipment and Reagent Requirements

From a T25 Flask (1 x 106 cells)

1. Transfer the cell contents from the T25 flask into a sterile 50

mL conical tube.

2. Centrifuge the 50 mL conical tube at 200 x g at 4℃ for 15

minutes.

3. Carefully remove all but approximately 100µL of the

supernatant using a pipette.

4. Gently resuspend the cell pellet in 12mL of fresh Cancer Stem

Premium™ medium (catalog no. 20101-100, or 20141-500),

pre-warmed to 37℃.

5. Aliquot 2mL of the cell suspension into each well of a single 6-

well plate (VWR, Cat. No. 29443-030).

6. Transfer the plate to a 37℃/5%CO2 cell culture incubator.

7. Add additional fresh Cancer Stem Premium™ medium at a

volume equivalent to 10% of the total culture volume every

two days to maintain the culture over a period of 7-10 days.

8. Tumor spheres will start to a form, and will be visible in

approximately 2-3 days, and after day 2 cells may be

passaged or used for experimentation.

Best wishes,

Ilya

Thanks Tsyrlow

Hello

yes this good protocol .

And see links and pdf .

Good Luck

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2708191/

http://www.promocell.com/fileadmin/promocell/PDF/Cancer_Stem_Cell_Medium.pdf