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脐带血血小板裂解物作为人骨髓间充质干细胞扩增中的血清替代物

Umbilical Cord Blood Platelet Lysate as Serum Substitute in Expansion of Human Mesenchymal Stem Cells.

作者信息

Shirzad Negin, Bordbar Sima, Goodarzi Alireza, Mohammad Monire, Khosravani Pardis, Sayahpour Froughazam, Baghaban Eslaminejad Mohamadreza, Ebrahimi Marzieh

机构信息

Department of Regenerative Biomedicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.

Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.

出版信息

Cell J. 2017 Oct;19(3):403-414. doi: 10.22074/cellj.2017.4886. Epub 2017 Aug 19.

Abstract

OBJECTIVES

The diverse clinical applications for human mesenchymal stem cells (hMSCs) in cellular therapy and regenerative medicine warrant increased focus on developing adequate culture supplements devoid of animal-derived products. In the present study, we have investigated the feasibility of umbilical cord blood-platelet lysate (UCB-PL) as a standard substitute for fetal bovine serum (FBS) and human peripheral blood-PL (PB-PL).

MATERIALS AND METHODS

In this experimental study, platelet concentrates (PC) from UCB and human PB donors were frozen, melted, and sterilized to obtain PL. Quality control included platelet cell counts, sterility testing (viral and microbial), total protein concentrations, growth factor levels, and PL stability. The effects of UCB-PL and PB-PL on hMSCs proliferation and differentiation into osteocytes, chondrocytes, and adipocytes were studied and the results compared with FBS.

RESULTS

UCB-PL contained high levels of protein content, platelet-derived growth factor- AB (PDGF-AB), and transforming growth factor (TGF) compared to PB-PL. All growth factors were stable for at least nine months post-storage at -70˚C. hMSCs proliferation enhanced following treatment with UCB-PL. With all three supplements, hMSCs could differentiate into all three lineages.

CONCLUSIONS

PB-PL and UCB-PL both were potent in hMSCs proliferation. However, PB promoted osteoblastic differentiation and UCB-PL induced chondrogenic differentiation. Because of availability, ease of use and feasible standardization of UCB-PL, we have suggested that UCB-PL be used as an alternative to FBS and PB-PL for the cultivation and expansion of hMSCs in cellular therapy.

摘要

目的

人间充质干细胞(hMSCs)在细胞治疗和再生医学中的多种临床应用,使得人们越来越关注开发不含动物源产品的合适培养补充剂。在本研究中,我们研究了脐带血血小板裂解液(UCB-PL)作为胎牛血清(FBS)和人外周血血小板裂解液(PB-PL)标准替代品的可行性。

材料与方法

在本实验研究中,将来自脐带血和人外周血供体的血小板浓缩物(PC)冷冻、融化并灭菌以获得血小板裂解液。质量控制包括血小板细胞计数、无菌检测(病毒和微生物)、总蛋白浓度、生长因子水平以及血小板裂解液稳定性。研究了UCB-PL和PB-PL对hMSCs增殖以及向骨细胞、软骨细胞和脂肪细胞分化的影响,并将结果与FBS进行比较。

结果

与PB-PL相比,UCB-PL含有高水平的蛋白质含量、血小板衍生生长因子AB(PDGF-AB)和转化生长因子(TGF)。所有生长因子在-70˚C储存后至少九个月内稳定。用UCB-PL处理后hMSCs增殖增强。使用所有三种补充剂时,hMSCs都可以分化为所有三个谱系。

结论

PB-PL和UCB-PL在hMSCs增殖方面都很有效。然而,PB促进成骨细胞分化,而UCB-PL诱导软骨形成分化。由于UCB-PL的可用性、易用性和可行的标准化,我们建议将UCB-PL用作细胞治疗中hMSCs培养和扩增的FBS和PB-PL替代品。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/5570405/aca0220db768/Cell-J-19-403-g01.jpg

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