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重复使用血液制品作为优化临床级脂肪来源间充质干细胞培养的替代补充物。

Re-using blood products as an alternative supplement in the optimisation of clinical-grade adipose-derived mesenchymal stem cell culture.

作者信息

Phetfong J, Tawonsawatruk T, Seenprachawong K, Srisarin A, Isarankura-Na-Ayudhya C, Supokawej A

机构信息

Centre for Research and Innovation, Mahidol University, Nakhon Pathom, Thailand.

Department of Orthopaedics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.

出版信息

Bone Joint Res. 2017 Jul;6(7):414-422. doi: 10.1302/2046-3758.67.BJR-2016-0342.R1.

Abstract

OBJECTIVES

Adipose-derived mesenchymal stem cells (ADMSCs) are a promising strategy for orthopaedic applications, particularly in bone repair. expansion of ADMSCs is required to obtain sufficient cell numbers. Xenogenic supplements should be avoided in order to minimise the risk of infections and immunological reactions. Human platelet lysate and human plasma may be an excellent material source for ADMSC expansion. In the present study, use of blood products after their recommended transfusion date to prepare human platelet lysate (HPL) and human plasma (Hplasma) was evaluated for culture expansion and osteogenesis of ADMSCs.

METHODS

Human ADMSCs were cultured in medium supplemented with HPL, Hplasma and a combination of HPL and Hplasma (HPL+Hplasma). Characteristics of these ADMSCs, including osteogenesis, were evaluated in comparison with those cultured in fetal bovine serum (FBS).

RESULTS

HPL and HPL+Hplasma had a significantly greater growth-promoting effect than FBS, while Hplasma exhibited a similar growth-promoting effect to that of FBS. ADMSCs cultured in HPL and/or Hplasma generated more colony-forming unit fibroblasts (CFU-F) than those cultured in FBS. After long-term culture, ADMSCs cultured in HPL and/or Hplasma showed reduced cellular senescence, retained typical cell phenotypes, and retained differentiation capacities into osteogenic and adipogenic lineages.

CONCLUSION

HPL and Hplasma prepared from blood products after their recommended transfusion date can be used as an alternative and effective source for large-scale expansion of ADMSCs. J. Phetfong, T. Tawonsawatruk, K. Seenprachawong, A. Srisarin, C. Isarankura-Na-Ayudhya, A. Supokawej. Re-using blood products as an alternative supplement in the optimisation of clinical-grade adipose-derived mesenchymal stem cell culture. Bone Joint Res 2017;6:414-422. DOI: 10.1302/2046-3758.67.BJR-2016-0342.R1.

摘要

目的

脂肪来源的间充质干细胞(ADMSC)是骨科应用中一种很有前景的策略,尤其在骨修复方面。需要扩增ADMSC以获得足够数量的细胞。应避免使用异种补充剂,以将感染和免疫反应的风险降至最低。人血小板裂解物和人血浆可能是ADMSC扩增的优质材料来源。在本研究中,评估了在推荐输血日期之后使用血液制品制备人血小板裂解物(HPL)和人血浆(Hplasma)用于ADMSC的培养扩增和成骨作用。

方法

将人ADMSC培养于添加了HPL、Hplasma以及HPL与Hplasma组合(HPL + Hplasma)的培养基中。与在胎牛血清(FBS)中培养的ADMSC相比,评估这些ADMSC的特性,包括成骨作用。

结果

HPL和HPL + Hplasma的促生长作用显著大于FBS,而Hplasma表现出与FBS相似的促生长作用。在HPL和/或Hplasma中培养的ADMSC比在FBS中培养的产生更多的集落形成单位成纤维细胞(CFU - F)。长期培养后,在HPL和/或Hplasma中培养的ADMSC显示细胞衰老减少,保留典型细胞表型,并保留向成骨和成脂谱系的分化能力。

结论

在推荐输血日期之后从血液制品制备的HPL和Hplasma可作为ADMSC大规模扩增的替代有效来源。J. Phetfong, T. Tawonsawatruk, K. Seenprachawong, A. Srisarin, C. Isarankura - Na - Ayudhya, A. Supokawej。重新利用血液制品作为优化临床级脂肪来源间充质干细胞培养的替代补充剂。《骨关节研究》2017;6:414 - 422。DOI:10.1302/2046 - 3758.67.BJR - 2016 - 0342.R1。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f4c5/5539302/b7c443aeb731/bonejointres-06-414-g001.jpg

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