Comparison study showing the growth and expansion of adipose-derived hMSCs using a variety of media, supplements, and attachment substrates

As more human mesenchymal stem/stromal cell (hMSC)-based therapies move closer to the clinic, the culture conditions used to propagate these cells in vitro are increasingly coming into the spotlight. We see this reflected every day by the growing number of labs now transitioning their standard culture systems to an environment that better supports the translational goals of their research.

Specifically, researchers are seeking cell culture reagents that are produced under cGMP guidelines, consistent from lot-to-lot, and able to meet the strict FDA standards for cell-based therapies, such as having a specific Drug Master File (DMF) and/or a history of successful use in clinical trials. Therefore, many MSC labs are now giving their cell culture an upgrade by replacing undefined, animal-derived, and variable components, such as fetal bovine serum (FBS), with serum- and animal component-free alternatives.

In this post, we are sharing the results of a thorough evaluation of both classical and serum-free MSC culture media performed by a prominent research team in California who reached out to Biological Industries USA to make this switch.

In the past, this lab routinely cultured adipose-derived hMSCs in a medium composed of alpha-MEM, L-glutamine, and FBS. However, based on their need to move their cell culture to a more clinically compliant system, this lab decided to evaluate clinical-grade reagents and alternatives to FBS – including the MSC NutriStem® culture system and PLTMax® Human Platelet Lysate.

For this evaluation, the lab compared the growth and expansion of adipose-derived hMSCs using a variety of basal media, supplements, and attachment substrates. The two basal media used were the lab’s standard alpha-MEM with 1% glutagro®, and the optimized MSC NutriStem® Basal Medium. Each of these basal media were then combined with 3 different supplements: 10% FBS, 5% PLTMax (noted in data as HPL), or 0.6% MSC NutriStem XF Supplement Mix (noted in data as XF). From there, cells in these 6 media combinations were cultured on 2 different attachment coatings, either BioCoat™ Fibronectin, or the MSC Attachment Solution, which is also a fibronectin matrix.

Overall, the lab found that the MSC NutriStem Basal Medium supplemented with PLTMax greatly outperformed all of the tested media combinations that included FBS for hMSC growth and expansion, demonstrating the effective use of PLTMax as an FBS alternative, even when supplemented at half the concentration (5% PLTMax vs 10% FBS). Additionally, the MSC NutriStem Basal Medium outperformed the use of alpha-MEM as a basal medium when supplemented with either PLTMax or the MSC NutriStem XF Supplement, showing a clear advantage to using a basal medium that’s been carefully optimized for hMSC culture.

Media Comparisons – Adipose Derived MSCs

Figure 1: Adipose-derived hMSCs cultured on a BioCoat™ fibronectin surface. Images represent cell expansion over 125 total hours, with media changes occurring at the 28- and 72-hour time-points. In this figure, “MSC NutriStem” refers to MSC NutriStem® Basal Medium, “HPL” refers to PLTMax, and “XF” refers to the MSC NutriStem® XF Supplement Mix.

Figure 2: Adipose-derived hMSCs cultured on wells coated with the MSC Attachment Solution. Images represent cell expansion over 125 total hours, with media changes occurring at the 28- and 72-hour time-points. In this figure, “MSC NutriStem” refers to MSC NutriStem® Basal Medium, “HPL” refers to PLTMax, and “XF” refers to the MSC NutriStem® XF Supplement Mix.

Human platelet lysate exhibits an effective alternative to FBS for hMSC culture

Together, these results show that human platelet lysates, such as PLTMax® and the new PLTGold®, make an effective animal serum-free alternative to FBS for hMSC culture, and can promote greater cell health and proliferation compared to FBS, particularly when combined with MSC NutriStem Medium.

The higher cell yields seen using PLTMax, PLTGold, and the MSC NutriStem culture system, combined with the fact that these reagents are all produced under cGMP and come with a long history of use in clinical trials, makes them a clear choice for many labs looking to move their cell culture one step closer to therapeutic use.

 

Would you like to evaluate the MSC NutriStem culture system, PLTMax, or PLTGold in your lab? Contact us at techsupport@bioindusa.com to learn more.

For more information on the reagents in this evaluation, click the links below, watch our on-demand webinar, or download our scientific poster.