Mesenchymal Stem Cell Culture Under SF, XF Conditions - Useful Tips

Mesenchymal Stem Cell Initial Isolation from Tissue

• For the initial isolation of human mesenchymal stem cells (hMSC) from tissue, add 2.5% of human AB serum to MSC NutriStem® XF.
• For the attachment and spreading of hMSC in serum-free conditions, it is highly recommended to use attachment factors (i.e. NutriCoat™ Attachment Solution; MSC Attachment Solution), specially treated TC-cultureware (i.e. CellBind®) or adherence promoting supplements (i.e. Platelet Lysate).

Seeding of Mesenchymal Stem Cells

• Recommended seeding concentration: 5000-6000cells/cm².
• Directly after seeding, tilt carefully the culture plate for achieving homogeneous of attached cells in the plate (under SF condition, the hMSC are rapidly attached, even after 5 min).
• Avoid using antibiotics.

Cell Passage

• Subculture when the culture reaches up to 80% confluence (higher confluence may induce cells differentiation, low viability).
• Use recommended dissociation solution, such as Recombinant Trypsin Solution.
• Under SF condition dilute the enzymatic solution using the complete MSC NutriStem® XF. Alternatively, use SBTI (Soy Bean Trypsin Inhibitor) for neutralization of trypsin.
• The dissociation time may differ mainly based on the quality of the cells (Older cells, confluent culture, and suboptimal culture conditions may lead to longer incubation time with dissociation solution).