Serum-converted platelet lysate can substitute for fetal bovine serum in human mesenchymal stromal cell cultures

被引:75
|
作者
Mojica-Henshaw, Mariluz P. [1 ]
Jacobson, Pam [1 ]
Morris, Julie [1 ]
Kelley, Linda [2 ]
Pierce, Jan [1 ]
Boyer, Michael [1 ]
Reems, Jo-Anna [1 ]
机构
[1] Univ Utah, Cell Therapy & Regenerat Med Facil, Salt Lake City, UT 84108 USA
[2] Cryo Cell Int Inc, Oldsmar, FL USA
关键词
blood platelets; cell proliferation; growth factors; mesenchymal stromal cells; platelet-rich plasma; HUMAN AB SERUM; STEM-CELLS; ANIMAL SERUM; MONONUCLEAR-CELLS; CALF SERUM; EXPANSION; HEPARIN; GROWTH; PROLIFERATION; DIFFERENTIATION;
D O I
10.1016/j.jcyt.2013.06.014
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Background aims. Fetal bovine serum (PBS) is commonly used as a serum supplement for culturing human mesenchymal stromal cells (hMSCs). However, human cells grown in FBS, especially for extended periods, risk potential exposure to bovine immunogenic proteins and infectious agents. To address this issue, we investigated the ability of a novel human platelet serum supplement to substitute for FBS in hMSC cultures. Methods. Platelet lysate-serum (PL-serum) was converted from platelet lysate-plasma (PL-plasma) that was manufactured from pooled platelet-rich plasma (PRP) apheresis units. Growth factor levels and the number of residual intact platelets in PL-serum and PL-plasma were compared with enzyme-linked immunosorbent assays and flow cytometry, respectively. Proliferation responses of hMSCs cultured in PL-serum, PL-plasma, or PBS were assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, the immunophenotype of harvested hMSCs was evaluated by flow cytometry and tri-lineage differentiation potential was evaluated by assessing adipogenic, osteogenic and chondrogenic development. Results. Selected growth factor levels in PL-serum were not significantly different from PL-plasma (P > 0.05). hMSC cultures supplemented with PL-serum had comparable growth kinetics to PL-plasma, and hMSC yields were consistently greater than with FBS. hMSCs harvested from cultures supplemented with PL-serum, PL-plasma or FBS had similar cell surface phenotypes and maintained tri-lineage differentiation potential. Conclusions. PL-serum, similar to PL-plasma, can substitute for PBS in hMSC cultures. Use of PL-serum, in contrast to PL-plasma, has an added advantage of not requiring addition of a xenogeneic source of heparin, providing a completely xeno-free culture medium.
引用
收藏
页码:1458 / 1468
页数:11
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