Human platelet lysate successfully promotes proliferation and subsequent chondrogenic differentiation of adipose-derived stem cells: a comparison with articular chondrocytes

Abstract

Fetal calf serum (FCS) bears a potential risk for carrying diseases and eliciting immune reactions. Nevertheless, it still represents the gold standard as medium supplement in cell culture. In the present study, human platelet lysate (PL) was tested as an alternative to FCS for the expansion and subsequent chondrogenic differentiation of human adipose-derived stem cells (ASCs). ASCs were expanded with 10% FCS (group F) or 5% PL (group P). Subsequently, three-dimensional (3D) micromass pellets were created and cultured for 5 weeks in chondrogenic differentiation medium. Additionally, the de- and redifferentiation potential of human articular chondrocytes (HACs) was evaluated and compared to ASCs. Both HACs and ASCs cultured with PL showed strongly enhanced proliferation rates. Redifferentiation of HACs was possible for cells expanded up to 3.3 population doublings (PD). At this stage, PL-expanded HACs demonstrated better redifferentiation potential than FCS-expanded cells. ASCs could also be differentiated following extended passaging. Glycosaminoglycan (GAG) quantification and qRT-PCR of 10 cartilage related markers demonstrated a tendency for increased chondrogenic differentiation of PL-expanded ASCs compared to cells expanded with FCS. Histologically, collagen type II but also collagen type X was mainly present in group P. The present study demonstrates that PL strongly induces proliferation of ASCs, while the chondrogenic differentiation potential is retained. HACs also showed enhanced proliferation and even better redifferentiation when previously expanded with PL. This suggests that PL is superior to FCS as a supplement for the expansion of ASCs and HACs, particularly with regard to chondrogenic (re)differentiation.

Partnership between platelet-rich plasma and mesenchymal stem cells: in vitro experience

“…Aiming to develop articular therapies by combining PRP and MSCs, Mishra et al 44 showed that buffered PRP enhanced the expression of chondrogenic markers, such as Sox9 and Aggrecan. These findings were corroborated in further studies 7,8,12,27,29,47 that showed enhanced chondrogenic differentiation, and extracellular cartilage matrix synthesis in the presence of PRP. Likewise, Shih et al 12 also proved that chondrogenic differentiation was enhanced and maintained through cell passages in PRP cultures comparing to FCS cultures.…”

“…After excluding reviews and articles pertaining to dental and bone biology, and adding hand search relevant articles, we included 57 articles as relevant to research in biological mechanisms, of which 22 articles perform large scale MSC production. Twelve articles perform studies on Adipose Stem Cells (ADSCs) [7][8][9][10][11][12][13][14][15][16][17][18] …”

Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy

“…Doucet et al (2005) describe the preparation and application of human platelet lysate (HPL) for expanding mesenchymal stem cells. Further studies show both autologous and allogeneic HPL to be superior to FBS in enhancing cell proliferation (Burnouf et al, 2016;Hildner et al, 2015;Hofbauer et al, 2014;Kim et al, 2015;Trojahn Kølle et al, 2013). HPL eliminates the risk of cross-species contamination, solves standing ethical considerations and meets the GMP guidelines.…”

A Luciferase-Based Quick Potency Assay to Predict Chondrogenic Differentiation

“…According to the analyses, the threshold for Col2A1 expression on mRNA level was set to 10 and together with the histological and immunohistological data we identified 5 good donors (3,4,5,8,9) and 4 bad donors (1, 2, 6, 7).…”

“…In contrast to bone marrow, adipose tissue can be harvested in large amounts by simple surgical procedure. However, a large donor variability for the chondrogenic differentiation potential has been reported 7,8 , which raises the risk for transplantation of low-potent cells. Therefore, a quick potency assay is required to predict cell potency before transplantation or long-term storage.…”