Isolation method and xeno-free culture conditions influence multipotent differentiation capacity of human Wharton’s jelly-derived mesenchymal stem cells

Stem Cell Research & Therapy

Marilena Lupu Plesu¹  Ilinca Gussi³  Livia Elena Sima²  Anca Remes¹  Mirel Adrian Popa¹  Maria Cristina Corotchi¹ 

[1] Angiogenesis and Cardiovascular Remodeling Group, Department of Regenerative Medicine, Institute of Cellular Biology and Pathology “Nicolae Simionescu” of the Romanian Academy, 8 B.P. Hasdeu Str., 050568, Bucharest, Romania;Department of Molecular Cell Biology, Institute of Biochemistry of the Romanian Academy, 296 Splaiul Independentei, 060031, Bucharest, Romania;Department of Obstetrics and Gynecology, Clinical Hospital “Dr. Ioan Cantacuzino”, 5-7 Ion Movila Str., 020000, Bucharest, Romania

关键词: Endothelial differentiation;    Plasticity;    Xeno-free medium;    Mesenchymal stem cells;    Wharton’s jelly;   

Others  :  847304
DOI  :  10.1186/scrt232

 received in 2013-03-11, accepted in 2013-07-08,  发布年份 2013

【 摘 要 】

Introduction

Human Wharton’s jelly (WJ) has become a preferred source of mesenchymal stem cells (MSCs) whose clinical applications are limited by the use of adequate xeno-free (XF), in vitro manipulation conditions. Therefore, the objective of our study was to characterize WJ-derived MSCs (WJ-MSCs), isolated by different methods and cultured in a commercially available, MSC XF medium, not least of all by investigating their endothelial differentiation capacity.

Methods

WJ explants and enzymatically dissociated WJ cells were cultured in a defined, XF medium for MSCs. Adherent cells at passages 2 and 5 were characterized as MSCs by flow cytometry, MTT, real-time quantitative reverse transcription PCR, and functional multipotent differentiation assays. The endothelial differentiation capacity of MSCs isolated and expanded until passage 2 in the MSC XF medium, and then subcultured for five passages in a commercially available endothelial growth medium (group A), was assessed over serial passages, as compared to adherent WJ-derived cells isolated and expanded for five consecutive passages in the endothelial medium (group B).

Results

The MSC phenotype of WJ explant- and pellet-derived cells, isolated and expanded in the MSC XF medium, was proven based on the expression of CD44/CD73/CD90/CD105 surface markers and osteo-/adipo-/chondrogenic multipotent differentiation potential, which differed according to the isolation method and/or passage number. Upon exposure to endothelial differentiation cues, cells belonging to group A did not exhibit endothelial cell characteristics over serial passages; by contrast, WJ pellet-derived cells belonging to group B expressed endothelial characteristics at gene, protein and functional levels, potentially due to culture conditions favoring the isolation of other stem/progenitor cell types than MSCs, able to give rise to an endothelial progeny.

Conclusions

The use of defined, MSC XF media for isolation and expansion of human WJ-MSCs is a prerequisite for the establishment of their real endothelial differentiation capacity, as candidates for clinical therapy applications. Thus, the standardization of WJ-MSCs isolation and culture expansion techniques in defined, MSC XF media, for their accurate characterization, would be a priority in the stem cell research field.

【 授权许可】

   
2013 Corotchi et al.; licensee BioMed Central Ltd.

【 图 表 】

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