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.
  • Always (even if serum is added) use MSC Attachment Solution for mesenchymal stem cell attachment and spreading.

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

  • Sub culture when 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).

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