Knockout Serum Replacement Accompanied With Basic Fibroblast Growth Factor in Culture Medium Promotes Growth of Human Breast Milk-Derived Cells.

AIM: Human breast milk is known to contain a variety of bioactive components, including stem and progenitor cells; however, their in vitro propagation is often constrained by limited adherence and yield. The use of conventional serum supplementation raises concerns regarding unnatural interactions and potential pathogen exposure. This study aims to assess the effects of serum and growth factors on the adhesion, proliferation, and propagation of human breast milk-derived cells (hBMCs).

METHODS

We cultured cells in four media: M1 (DMEM + 10% FBS), M2 (DMEM/F12 + 20% KnockOut Serum Replacement [KSR] + 10 ng/mL bFGF), M3 (DMEM/F12 + 10% KSR + 10 ng/mL bFGF), and M4 (DMEM + 10% FBS + 5% HS). Cell adhesion, proliferation, and morphology were evaluated using bright-field microscopy, fluorescence imaging, and cell viability assays.

RESULTS

Our findings indicated postpartum-dependent variations in hBMC numbers and demonstrated that serum-free conditions with KSR significantly enhanced cell adhesion and proliferation. Specifically, M2 and M3 facilitated increased proliferation with reduced doubling time, resulting in higher cell yield. Furthermore, the study underscored the impact of contamination and fat globules on lower adhesion efficiency.

CONCLUSIONS

Overall, these results identify KSR-based media as a promising alternative to FBS, providing optimized conditions for the propagation of hBMCs and paving the way for their future therapeutic and experimental applications.