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外周血和脐带血血小板裂解物在多能间充质基质细胞分离和扩增中的评估

Evaluation of Peripheral Blood and Cord Blood Platelet Lysates in Isolation and Expansion of Multipotent Mesenchymal Stromal Cells.

作者信息

Christou Ioanna, Mallis Panagiotis, Michalopoulos Efstathios, Chatzistamatiou Theofanis, Mermelekas George, Zoidakis Jerome, Vlahou Antonia, Stavropoulos-Giokas Catherine

机构信息

Hellenic Cord Blood Bank, Biomedical Research Foundation Academy of Athens, 4 Soranou Ephessiou Street, 115 27 Athens, Greece.

Biotechnology division, Biomedical Research Foundation Academy of Athens, 4 Soranou Ephessiou Street, 115 27 Athens, Greece.

出版信息

Bioengineering (Basel). 2018 Feb 26;5(1):19. doi: 10.3390/bioengineering5010019.

Abstract

: Multipotent Mesenchymal Stromal Cells (MSCs) are used in tissue engineering and regenerative medicine. The in vitro isolation and expansion of MSCs involve the use of foetal bovine serum (FBS). However, many concerns have been raised regarding the safety of this product. In this study, alternative additives derived either from peripheral or cord blood were tested as an FBS replacement. Platelet lysates (PL) from peripheral and cord blood were used for the expansion of MSCs. The levels of growth factors in peripheral blood (PB) and cord blood (CB) PLs were determined using the Multiple Reaction Monitoring (MRM). Finally, the cell doubling time (CDT), tri-lineage differentiation and phenotypic characterization of the MSCs expanded with FBS and PLs were determined. MSCs treated with culture media containing FBS and PB-PL, were successfully isolated and expanded, whereas MSCs treated with CB-PL could not be maintained in culture. Furthermore, the MRM analysis yielded differences in growth factor levels between PB-PL and CB-PL. In addition, the MSCs were successfully expanded with FBS and PB-PL and exhibited tri-lineage differentiation and stable phenotypic characteristics. PB-PL could be used as an alternative additive for the production of MSCs culture medium applied to xenogeneic-free expansion and maintenance of MSCs in large scale clinical studies.

摘要

多能间充质基质细胞(MSCs)被用于组织工程和再生医学。MSCs的体外分离和扩增涉及使用胎牛血清(FBS)。然而,人们对该产品的安全性提出了许多担忧。在本研究中,测试了源自外周血或脐带血的替代添加剂作为FBS的替代品。来自外周血和脐带血的血小板裂解物(PL)用于MSCs的扩增。使用多反应监测(MRM)测定外周血(PB)和脐带血(CB)PL中生长因子的水平。最后,测定了用FBS和PLs扩增的MSCs的细胞倍增时间(CDT)、三系分化和表型特征。用含有FBS和PB-PL的培养基处理的MSCs成功分离并扩增,而用CB-PL处理的MSCs不能在培养中维持。此外,MRM分析显示PB-PL和CB-PL之间生长因子水平存在差异。此外,MSCs用FBS和PB-PL成功扩增,并表现出三系分化和稳定的表型特征。PB-PL可作为一种替代添加剂,用于生产MSCs培养基以应用于无异种的大规模临床研究中MSCs的扩增和维持。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b820/5874885/508c6f66c422/bioengineering-05-00019-g001.jpg

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