Pathogen-free, plasma-poor platelet lysate and expansion of human mesenchymal stem cells

Journal of Translational Medicine

Luca Pierelli²  Marianna Nuti²  Luigi Frati²  Pierangela Totta¹  Claudio Lavorino³  Michelina Miceli³  Giuseppina Bonanno⁴  Daniela Fioravanti³  Paola Iudicone³ 

[1] Department of Science, Section Biomedical Science and Technology, University “Roma Tre”, Rome, Italy;Department of Experimental Medicine, Sapienza University, Rome, Italy;Immunohematology and Transfusion Medicine, San Camillo Forlanini Hospital, Rome, Italy;Department of Gynecology, Catholic University Medical School, Rome, Italy

关键词: Platelet lysate;    Mesenchymal stem cells;    Cellular therapy;    Blood component preparations;   

Others  :  821811
DOI  :  10.1186/1479-5876-12-28

 received in 2013-10-04, accepted in 2014-01-23,  发布年份 2014

【 摘 要 】

Background

Supplements to support clinical-grade cultures of mesenchymal stem cells (MSC) are required to promote growth and expansion of these cells. Platelet lysate (PL) is a human blood component which may replace animal serum in MSC cultures being rich in various growth factors. Here, we describe a plasma poor pathogen-free platelet lysate obtained by pooling 12 platelet (PLT) units, to produce a standardized and safe supplement for clinical-grade expansion of MSC.

Methods

PL lots were obtained by combining 2 6-unit PLT pools in additive solution (AS) following a transfusional-based procedure including pathogen inactivation (PI) by Intercept technology and 3 cycles of freezing/thawing, followed by membrane removal. Three PI-PL and 3 control PL lots were produced to compare their ability to sustain bone marrow derived MSC selection and expansion. Moreover, two further PL, subjected to PI or not, were also produced starting from the same initial PLT pools to evaluate the impact of PI on growth factor concentration and capacity to sustain cell growth. Additional PI-PL lots were used for comparison with fetal bovine serum (FBS) on MSC expansion. Immunoregulatory properties of PI-PL-generated MSC were documented in vitro by mixed lymphocyte culture (MLC) and peripheral blood mononuclear cells (PBMC) mitogen induced proliferation.

Results

PI-PL and PL control lots had similar concentrations of 4 well-described growth factors endowed with MSC stimulating ability. Initial growth and MSC expansion by PI-PL and PL controls were comparable either using different MSC populations or in head to head experiments. Moreover, PI-PL and PL control sustained similar MSC growth of frozen/thawed MSC. Multilineage differentiation of PI-derived and PI-PL-derived MSC were maintained in any MSC cultures as well as their immunoregulatory properties. Finally, no direct impact of PI on growth factor concentration and MSC growth support was observed, whereas the capacity of FBS to sustain MSC expansion in basic medium was irrelevant as compared to PL and PI-PL.

Conclusion

The replacement of animal additives with human supplements is a basic issue in MSC ex vivo production. PI-PL represents a standardized, plasma-poor, human preparation which appears as a safe and good candidate to stimulate MSC growth in clinical-scale cultures.

【 授权许可】

   
2014 Iudicone et al.; licensee BioMed Central Ltd.

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