Question

1. Assume that you should prepare medium with DMEM containing 10 % FBS, 150U/mL Penicillin, 150μg/mL Streptomycin and 1,5 mM L-glutamine for Human Mesenchymal Stem Cells (hMSC) cell line cultivation. (20 pts)

Penicillin Stock : 10,000 U/mL

Streptomycin Stock: 10mg/mL

Molecular Weight of L-Glutamine: 146,14 g/mol

HINT 1: L-glutamine is powder. 10ml stock solution should be prepared in PBS by dissolving desired amount of L-glutamine.

HINT 2: The volume of L-glutamine should be 1% of total medium volume.

Please find required

1. FBS volume
2. Penicillin volume
3. Streptomycin volume
4. L-Glutamine volume
5. L- Glutamine amount in the stock solution

to prepare 50 ml basal medium according to stock solution concentration.

Answer 1

Total volume to be prepared is 50ml.

The volume of

1. FBS to be prepared = 10% = 10×50/100 = 5ml.

2. Penicillin volume: 10000U in 1000ul, then 150U in ?ul is required and hence, 150× 1000/ 10000 = 15ul. Then for 50ml, volume required is 15×50=750ul.

3. Streptomycin volume: 10000ug in 1000ul then 150ug in ?ul. = 1000×150/10000 = 15ul. Then for 50ml, volume required is 15×50= 750ul.

4 and 5. To make 1.5mM L-glutamine solution in 10ml stock solution,

1.5mM = wt×1000/14614×10

Wt. in 10ml stock solution = 219.21mg

Volume required to prepare 50ml basal medium is = 0.5ml stock solution in 50ml.

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