Cell proliferation of human bone marrow mesenchymal stem cells on biodegradable microcarriers enhancesin vitrodifferentiation potential

Abstract

Objectives: For reasons of provision of highlyspecific surface area and three-dimensional culture, microcarrier culture (MC) has garnered great interest for its potential to expand anchorage-dependent stem cells. This study utilizes MC for in vitro expansion of human bone marrow mesenchymal stem cells (BMMSCs) and analyses its effects on BMMSC proliferation and differentiation. Materials and methods: Effects of semi-continuous MC compared to control plate culture (PC) and serial bead-to-bead transfer MC (MC bead-T) on human BMMSCs were investigated. Cell population growth kinetics, cell phenotypes and differentiation potential of cells were assayed. Results: Maximum cell density and overall fold increase in cell population growth were similar between PCs and MCs with similar starting conditions, but lag period of BMMSC growth differed substantially between the two; moreover, MC cells exhibited reduced granularity and higher CXCR4 expression. Differentiation of BMMSCs into osteogenic and adipogenic lineages was enhanced after 3 days in MC. However, MC bead-T resulted in changes in cell granularity and lower osteogenic and adipogenic differentiation potential. Conclusions: In comparison to PC, MC supported expansion of BMMSCs in an up-scalable threedimensional culture system using a semi-continuous process, increasing potential for stem cell homing ability and osteogenic and adipogenic differentiation.

hMSC Production in Disposable Bioreactors with Regards to GMP and PAT

“…Regarding a constant final cell density at the end of the process, the inoculation density has an influence on the process efficiency as the expansion factor is influenced. Therefore, the lowest inoculation density (≈ 0.6 · 10 3 cells/cm 2 ) achieved the highest expansion factor (≈ 7-fold) [25,30].…”

Culture of human mesenchymal stem cells on microcarriers in a 5 l stirred-tank bioreactor

“…Cultispher-S (Eibes et al 2010), Cultispher-G (Sun et al 2010) and SoloHill Plastic P102-L (Dos Santos et al 2011). However in all the aforementioned studies, the largest volume used was no more than 200 mL and it is unclear as to whether samples of microcarrier suspension greater than 2 mL were harvested.…”

Increased Differentiation Capacity of Bone Marrow-Derived Mesenchymal Stem Cells in Aquaporin-5 Deficiency

“…Lipid vesicles were stained by 2% (w/v) Oil red O (SigmaAldrich). The adipogenic differentiation potential of BMMSCs was further quantified by flow cytometry after staining of lipid vesicles with 0.5 mg/mL Nile Red dye (Sigma-Aldrich) as described previously [19].…”