Comparison of Human Platelet Lysate versus Fetal Bovine Serum for Expansion of Human Articular Cartilage–Derived Chondroprogenitors

Abstract

Purpose Articular chondroprogenitors, a suitable contender for cell-based therapy in cartilage repair, routinely employ fetal bovine serum (FBS) for expansion and differentiation. The possibility of transplant rejections or zoonoses transmissions raise a need for xeno-free alternatives. Use of human platelet lysate (hPL), a nutrient supplement abundant in growth factors, has not been reported for human chondroprogenitor expansion thus far. Our aim was to compare the biological profile of chondroprogenitors grown in hPL versus FBS. Methods Chondroprogenitors were isolated from 3 osteoarthritic knee joints. Following differential fibronectin adhesion assay, passage 0 cells grown in (a) 10% FBS and (b) 10% hPL were considered for assessment of growth kinetics, surface marker expression, gene expression, and trilineage differentiation. Latent transforming growth factor–β1 (TGFβ1) levels were also measured for each culture medium used. Results Cellular proliferation was significantly higher in cells grown with hPL ( P < 0.01). Surface marker expression was comparable except in CD-146 where hPL group had significantly higher values ( P = 0.03). Comparison of mRNA expression revealed notably low values of collagen I, collagen X, aggrecan, and collagen II ( P < 0.05). Trilineage differentiation was seen in both groups with higher alizarin red uptake noted in hPL. There were also significantly higher levels of latent TGFβ1 in the medium containing hPL as compared to FBS. Conclusions This is the first in vitro xeno-free study to affirm that hPL can serve as an optimal growth supplement for expansion of articular chondroprogenitors, although an in-depth assessment of resident growth factors and evaluation of different dilutions of hPL is required to assess suitability for use in translational research.

Pondering the Potential of Hyaline Cartilage–Derived Chondroprogenitors for Tissue Regeneration: A Systematic Review

“…14 Supplementation with platelet lysate as opposed to FBS caused chondroprogenitors to show comparatively higher alizarin red uptake. 17 Commonly reported positive MSC markers included CD105 (n = 17), CD90 (n = 20), CD73 (n = 12), while negative MSC markers included CD34 (n = 15) and CD45 (n = 15). Numerous reports evaluated the following cell surface proteins as potential markers of chondrogenesis namely CD29 (n = 12) in combination with CD49e (n = 19), CD146 (n = 4), and CD166 (n = 13).…”

“…14 It was also seen that for creation of xeno-free culture conditions, when human platelet lysate was used in place of FBS, progenitors displayed greater proliferation with lower expression of type I and X collagen (fibrocartilage and hypertrophy markers). 17 With regard to culture conditions producing maximum growth of progenitors, Melero et al report an optimal seeding density of 10,000 cells/cm 2 and medium volume of 0.3 mL/cm 2 . 18,19 The authors further demonstrated an efficient post-cryoviability and 17-fold expansion potential using macroporous microcarrier supplemented with 40% FCS and 1 ng/mL of TGFβ1.…”

Feasibility of Human Platelet Lysate as an Alternative to Foetal Bovine Serum for In Vitro Expansion of Chondrocytes

“…In addition, Muraglia et al also reported that addition of HPL to FBS supplement significantly enhanced the chondrocyte proliferation [42]. Kachroo et al compared the expansion of chondroprogenitor cells isolated from cartilage with HPL and FBS [43]. The authors found that chondroprogenitor cells cultured with HPL have higher proliferation but lower expression of chondrogenic (aggrecan and col II), dedifferentiation (col I) and hypertrophic markers (col X) compared to those expanded with FBS.…”

Migratory chondroprogenitors retain superior intrinsic chondrogenic potential for regenerative cartilage repair as compared to human fibronectin derived chondroprogenitors

“…Another standard method to obtain progenitors is based on their migratory potential from cartilage explants in culture 8 , 18 . Both fibronectin adhesion assay derived 19 , 20 and migratory chondroprogenitors 8 , 21 have been likened to MSCs as delineated by the International Society for Cellular Therapy (ISCT) 2006 22 , demonstrating plastic adherence, similar surface marker expression, and trilineage differentiation potential. Extensive work on these progenitors has provided information on their growth characteristics, where fibronectin-derived cells displayed clonal growth and required additional growth factors for expansion, unlike migratory progenitors 23 .…”