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

Abstract

ObjectivesAdipose-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.MethodsHuman 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).ResultsHPL 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.ConclusionHPL 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.

Human platelet lysate to substitute fetal bovine serum in hMSC expansion for translational applications: a systematic review

“…In the vast majority of reports the ability of hMSCs to be differentiated along pre-determined lineages was not affected by being cultured using hPL: adipogenic, osteogenic and chondrogenic differentiation were demonstrated with no relevant difference in effectiveness or potential to the same hMSCs cultured with FBS. In three cases MSCs, expanded in hPL, were more effectively differentiated along osteogenic, chondrogenic and adipogenic routes [ 37 , 49 , 58 ]; others reported a difference only for osteogenic [ 17 , 40 , 42 , 47 , 54 ] or adipogenic differentiation respectively [ 19 , 31 ]. Only one publication reported that FBS supported osteogenic and adipogenic differentiation to a higher degree [ 57 ].…”

Adipose Tissue Stem Cells for Therapy: An Update on the Progress of Isolation, Culture, Storage, and Clinical Application

“…Besides, human serum enhances the ASCs proliferation without any differences in morphology or immunostaining profile compared to traditional culture with FBS [53]. Blood-derived products consisted of many growth factors compared to FBS or FCS can be useful to maintain the high proliferation rate, stem cell properties, and differentiation characteristics of ASCs in long-term cultivation [53,56]. Moreover, the combination of platelet lysate and human plasma which are isolated from expired blood can reduce senescence of ASCs during cultivation [56].…”

“…Blood-derived products consisted of many growth factors compared to FBS or FCS can be useful to maintain the high proliferation rate, stem cell properties, and differentiation characteristics of ASCs in long-term cultivation [53,56]. Moreover, the combination of platelet lysate and human plasma which are isolated from expired blood can reduce senescence of ASCs during cultivation [56]. Besides, platelet-rich plasma added medium can stimulate the differentiation of ASCs into chondroblasts [57].…”

The Effect of Blood-Derived Products on the Chondrogenic and Osteogenic Differentiation Potential of Adipose-Derived Mesenchymal Stem Cells Originated from Three Different Locations

“…Preclinical data indicate that blood-derived products support the maintenance of cell proliferation, as well as stem cell properties and differentiation characteristics in in vitro settings [26,27]. Moreover, a combination of platelet lysate and human plasma may reduce senescence of AD-MSCs in culture [27]. PRP-enriched medium specifically was shown to enhance chondrogenic differentiation of AD-MSCs [28].…”