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

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

doi:10.1302/2046-3758.67.BJR-2016-0342.R1

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

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

Authors

J Phetfong¹, T Tawonsawatruk², K Seenprachawong³, A Srisarin³, C Isarankura-Na-Ayudhya⁴, A Supokawej⁵

Affiliations

¹ Centre for Research and Innovation, Mahidol University, Nakhon Pathom, Thailand.
² Department of Orthopaedics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
³ Department of Clinical Microscopy, Mahidol University, Nakhon Pathom, Thailand.
⁴ Centre for Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand.
⁵ Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University, 999 Phutthamonthon 4 Road, Salaya, Phuttamonthon, Nakhon Pathom 73170, Thailand aungkura.jer@mahidol.ac.th.

PMID: 28720606
PMCID: PMC5539302
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. Ex vivo 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 in vitro 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 ex vivo expansion of ADMSCs.Cite this article: 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.

Keywords: Adipose-derived mesenchymal stem cells; Human plasma; Mesenchymal stem cells; Platelet lysate; Stem cell culture.