Hi, Rezaie,

As for as serum preparation, you can follow the standard way

1.Adipose Mesenchymal Stromal Cells Isolated From Type 2 Diabetic Patients Display Reduced Fibrinolytic Activity.

2. The serum profile of adipokines in naïve patients with diabetes mellitus type 2 and obesity

3. A simple and fast liquid–liquid extraction method for the determination of 2,2′,4,4′,5,5′-hexachlorobiphenyl (CB-153) and 1,1-dichloro-2,2-bis(p-chlorophenyl)-ethylene (p,p′-DDE) from human serum for epidemiological studies on type 2 diabetes

For Exosomes, you can check the protocols mentioned in these papers:

1. Analysis of circulating microRNAs that are specifically increased in hyperlipidemic and/or hyperglycemic sera.

2. Serum microRNA profiling of hyperlipidemic and/or hyperglycemic patients

reveals specifically increased levels of miR-122, miR-125a, miR-486 and miR-92a

3. Analyzing the Circulating MicroRNAs in Exosomes/Extracellular Vesicles from Serum or Plasma by qRT-PCR

4. Noble Polymeric Surface Conjugated with Zwitterionic Moieties and Antibodies for the Isolation of Exosomes from Human Serum

5. Analysis of the RNA content of the exosomes derived from blood serum and urine and its potential as biomarkers

6. The majority of microRNAs detectable in serum and saliva is concentrated in exosomes

Dear Yasar

than you for your answer but  the main questions are : 1. can I use sera without filtration or heating instead of  FBS?

2.  Do sera Exosomes confound my test (MSC-derived Exosomes isolation)?

1. Preparation of sera from human blood can be found on the Web

    http://www.proimmune.com/ecommerce/pdf_files/PR31.pdf (2nd page)

    http://molmeth.org/preparation-serum-whole-blood

   After collecting serum from tube, you may have to heat inactivate just as of FBS.

2. Yes, sera contain exosomes. You can remove exosomes from sera by ultracentifugation (100,000 x g for >2 hr, mostly 130 min, 4 degree celcius).

Ref:

Dendritic Cell-Derived Exosomes Need To Activate Both T and B Cells To Induce Antitumor Immunity

Tanja I. Näslund, Ulf Gehrmann, Khaleda R. Qazi, Mikael C. I. Karlsson and Susanne Gabrielsson

http://www.jimmunol.org/content/190/6/2712

Dear Dr. soo Kim

Good guidance. Thank you for you helps  

Excuse me Please, but I have 2 questions:

 can I remove exosomes by ExoQuick kit from sera?

And after remove exosome from sera, will the cells grow better?

Form the questions you have added, I guess you don't want any residual exosomes in your sera. Am I right?

1. Can I remove exosomes by ExoQuick kit from sera?

You have to read the manufacturer's materials (https://www.systembio.com/downloads/Manual_ExoQuick_WEB.pdf) for the answer. I do not know what material(s) they use for spin down exosome from sera.

The material may or may not affect the exosome-depleted serum.

Although the answer for your question is "yes, you can remove exosome from sera. Yet, it is uncertain whehter you can use the exosome-depleted sera (that may contain chemicals from the Exoquick reagent) for your experiment. You need to consult with the manufacturer to make sure.

2. After remove exosome from sera, will the cells grow better?

On the same token, I don't have an answer for this. You have to talk or communicate with the manufacturer to find out whether  exosome-depleted serum can be used for biological studies including cell culture,

Thank you for your time and helps