The best media and serum replacement combination for adipose-derived stem cell culture

by Alexey Bersenev on February 18, 2016 · 0 comments

in adipose, mesenchymal

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There are a lot of differences between labs in media and supplements for growing of mesenchymal stromal cells (MSC). The best way to find your own best combination is to set up a comparative study. Unfortunately, such studies still rare find. Recently one of the most interesting comparative studies was published in Stem Cells TM. A group of researchers from Denmark compared 5 culture conditions for growth and characteristics of human MSCs, derived from adipose tissue. The following combinations were compared:

  1. StemPro alone (commercially available serum-free, xeno-free medium)
  2. DMEM + 10% PL (commercially available human platelet lysate -Stemulate)
  3. a-MEM + 10% Fetal Calf Serum (FCS)
  4. a-MEM + 5% PL
  5. a-MEM + 10% PL

Other additives were the same for all 5 tested conditions and included: Penicillin/ Strepromycin and GlutaMAX. MSC were isolated from adipose tissue-derived stromal vascular fraction of 5 normal donor, seeded in the same density and cultured in the same type of flask. Proliferation, attachment, colony-forming ability (CFU) and surface markers expression were compared.

Turns out that MSC growth and proliferation was better in all media with PL, compared to StemPro and FCS at 2-4 passages. StemPro did not cosistently support MSC culture.

PDL(Figure 1C from Riis, et al, 2016, DOI: 10.5966/sctm.2015-0148, modified)

StemPro supported cell attachment better than other media, DMEM was the worst. MSC, cultured in a-MEM with 5% and 10% of PL were larger, more granular and heterogenous. Frequency of CFU was significantly less in StemPro and not different between other conditions. As for expression of cell surface markers, no difference was observed between a-MEM 5% PL and 10% PL and DMEM 10% PL. Neither a-MEM vs. DMEM nor PL vs. FCS significantly impact immunophenotype. Expression of of CD73 and CD105 was more bright, but CD146 was dim in FCS medium, compared to PL media. MSC, cultured in a-MEM were more homogenous in terms of subpopulations. Expression of CD146 decreased with passage (from 1 to 7) in all conditions, except a-MEM 10% PL, where it was maintained at higher level.

The most important conclusions from the study:

We found, however, that StemPro, the chemically defined and xenogeneic-free medium used in this study, did not support the establishment of a viable culture of ASCs from SVF, although it was previously shown to satisfactorily support ASC expansion…
… culture in hPL favored a high proliferation rate, and both FCS and hPL were appropriate for maintaining stem cell characteristics. However, culture in either FCS or hPL favored specific subpopulations within the ASCs in terms of the intensity of specific surface antigen expression.

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