Due to the limited self-healing capacity of articular cartilage, innovative, regenerative approaches are of particular interest. The use of two-stage procedures utilizing in vitro-expanded mesenchymal stromal cells (MSCs) from various cell sources requires good manufacturing practice-compliant production, a process with high demands on time, staffing, and financial resources. In contrast, one- stage procedures are directly available, but need a safe enrichment of potent MSCs. CD271 is a surface marker known to marking the majority of native MSCs in bone marrow (BM). In this study, the feasibility of generating a single-stage cartilage graft of enriched CD271+ BM-derived mononuclear cells (MNCs) without in vitro monolayer expansion from eight healthy donors was investigated. Cartilage grafts were generated by magnetic-activated cell sorting and separated cells were directly transferred into collagen type I hydrogels, followed by 3D proliferation and differentiation period of CD271+, CD271−, or unseparated MNCs. CD271+ MNCs showed the highest proliferation rate, cell viability, sulfated glycosaminoglycan deposition, and cartilage marker expression compared to the CD271− or unseparated MNC fractions in 3D culture. Analysis according to the minimal criteria of the International Society for Cellular Therapy highlighted a 66.8-fold enrichment of fibroblast colony-forming units in CD271+ MNCs and the only fulfillment of the MSC marker profile compared to unseparated MNCs. In summary, CD271+ MNCs are capable of generating adequate articular cartilage grafts presenting high cell viability and notable chondrogenic matrix deposition in a CE-marked collagen type I hydrogel, which can obviate the need for an initial monolayer expansion.
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References
1.
FilardoG, MadryH, JelicM, RoffiA, CucchiariniM and KonE. (2013). Mesenchymal stem cells for the treatment of cartilage lesions: from preclinical findings to clinical application in orthopaedics. Knee Surg Sports Traumatol Arthrosc, 21:1717–1729.
2.
de GirolamoL, LucarelliE, AlessandriG, AvanziniMA, BernardoME, BiagiE, BriniAT, D'AmicoG, FagioliF, et al. (2013). Mesenchymal stem/stromal cells: a new “cells as drugs” paradigm. Efficacy and critical aspects in cell therapy. Curr Pharm Des, 19:2459–2473.
3.
GobbiA, ScottiC, KarnatzikosG, MudhigereA, CastroM and PerettiGM. (2017). One-step surgery with multipotent stem cells and Hyaluronan-based scaffold for the treatment of full-thickness chondral defects of the knee in patients older than 45 years. Knee Surg Sports Traumatol Arthrosc, 25:2494–2501.
4.
BornesTD, AdesidaAB and JomhaNM. (2014). Mesenchymal stem cells in the treatment of traumatic articular cartilage defects: a comprehensive review. Arthritis Res Ther, 16:432.
5.
TorreML, LucarelliE, GuidiS, FerrariM, AlessandriG, de GirolamoL, PessinaA and FerreroI. (2015). Ex vivo expanded mesenchymal stromal cell minimal quality requirements for clinical application. Stem Cells Dev, 24:677–685.
6.
The European Parliament and the Council of the European Union. (2007). Regulation (EC) No 1394/2007 of the European Parliament and of the Council of 13 November 2007 on Advanced Therapy Medicinal Products and Amending Directive 2001/83/EC and Regulation (EC) No 726/2004.
7.
SensebéL, KramperaM, SchrezenmeierH, BourinP and GiordanoR. (2010). Mesenchymal stem cells for clinical application. Vox Sang, 98:93–107.
8.
DahlJ-A, DuggalS, CoulstonN, MillarD, MelkiJ, ShahdadfarA, BrinchmannJE and CollasP. (2008). Genetic and epigenetic instability of human bone marrow mesenchymal stem cells expanded in autologous serum or fetal bovine serum. Int J Dev Biol, 52:1033–1042.
9.
EndresM, NeumannK, HauplT, ErggeletC, RingeJ, SittingerM and KapsC. (2007). Synovial fluid recruits human mesenchymal progenitors from subchondral spongious bone marrow. J Orthop Res, 25:1299–1307.
10.
JonesE and SchäferR. (2015). Biological differences between native and cultured mesenchymal stem cells: implications for therapies. Methods in molecular biology (Clifton, N.J.), 1235:105–120.
11.
ChahlaJ, DeanCS, MoatsheG, Pascual-GarridoC, Serra CruzR and LaPradeRF. (2016). Concentrated bone marrow aspirate for the treatment of chondral injuries and osteoarthritis of the knee: a systematic review of outcomes. Orthop J Sports Med, 4:2325967115625481.
12.
NiemeyerP, AlbrechtD, AndereyaS, AngeleP, AteschrangA, AurichM, BaumannM, BoschU, ErggeletC, et al. (2016). Autologous chondrocyte implantation (ACI) for cartilage defects of the knee: a guideline by the working group “Clinical Tissue Regeneration” of the German Society of Orthopaedics and Trauma (DGOU). Knee, 23:426–435.
13.
GracitelliGC, MoraesVY, FrancioziCE, LuzoMV and BellotiJC. (2016). Surgical interventions (microfracture, drilling, mosaicplasty, and allograft transplantation) for treating isolated cartilage defects of the knee in adults. Cochrane Database Syst Rev, 9:CD010675.
14.
CalloniR, Cordero, AparicioElvira Alicia, Henriques, PêgasJoão Antonio and BonattoD. (2013). Reviewing and updating the major molecular markers for stem cells. Stem Cells Dev, 22:1455–1476.
15.
KuçiS, KuçiZ, KreyenbergH, DeakE, PütschK, HueneckeS, AmaraC, KollerS, RettingerE, et al. (2010). CD271 antigen defines a subset of multipotent stromal cells with immunosuppressive and lymphohematopoietic engraftment-promoting properties. Haematologica, 95:651–659.
16.
QuiriciN, SoligoD, BossolascoP, ServidaF, LuminiC and DeliliersGL. (2002). Isolation of bone marrow mesenchymal stem cells by anti-nerve growth factor receptor antibodies. Exp Hematol, 30:783–791.
17.
GhazanfariR, LiH, ZacharakiD, LimHC and SchedingS. (2016). Human non-hematopoietic CD271pos/CD140alow/neg bone marrow stroma cells fulfill stringent stem cell criteria in serial transplantations. Stem Cells Dev, 25:1652–1658.
18.
LiH, GhazanfariR, ZacharakiD, LimHC and SchedingS. (2016). Isolation and characterization of primary bone marrow mesenchymal stromal cells. Ann N Y Acad Sci, 1370:109–118.
19.
MifuneY, MatsumotoT, MurasawaS, KawamotoA, KurodaR, ShojiT, KurodaT, FukuiT, KawakamiY, KurosakaM and AsaharaT. (2013). Therapeutic superiority for cartilage repair by CD271-positive marrow stromal cell transplantation. Cell Transplant, 22:1201–1211.
20.
Álvarez-ViejoM, Menéndez-MenéndezY and Otero-HernándezJ. (2015). CD271 as a marker to identify mesenchymal stem cells from diverse sources before culture. World J Stem Cells, 7:470–476.
21.
JonesE, EnglishA, ChurchmanSM, KouroupisD, BoxallSA, KinseyS, GiannoudisPG, EmeryP and McGonagleD. (2010). Large-scale extraction and characterization of CD271+ multipotential stromal cells from trabecular bone in health and osteoarthritis: implications for bone regeneration strategies based on uncultured or minimally cultured multipotential stromal cells. Arthritis Rheum, 62:1944–1954.
22.
RozemullerH, PrinsH-J, NaaijkensB, StaalJ, BühringH-J and MartensAC. (2010). Prospective isolation of mesenchymal stem cells from multiple mammalian species using cross-reacting anti-human monoclonal antibodies. Stem Cells Dev, 19:1911–1921.
23.
DominiciM, Le BlancK, MuellerI, Slaper-CortenbachI, MariniF, KrauseD, DeansR, KeatingA, ProckopD and HorwitzE. (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8:315–317.
24.
JahroomishiraziR, BaderA, EbertS, SchmidtC, SedaghatiB, Schulz-SiegmundM and ZscharnackM. (2014). Isolation and characterization of CD271(+) stem cells derived from sheep dermal skin. Cells Tissues Organs, 200:141–152.
25.
European Medicines Agency. (2013). MACI - matrix-applied characterised autologous cultured chondrocytes: Annex I Summary of Product Characteristics. Available from: www.ema.europa.eu/docs/en_GB/document_library/EPAR_-_Product_Information/human/002522/WC500145886.pdf (last accessed June13, 2016).
26.
PoloniA, MauriziG, RosiniV, MondiniE, ManciniS, DiscepoliG, BiasioS, BattagliniG, FelicettiS, et al. (2009). Selection of CD271(+) cells and human AB serum allows a large expansion of mesenchymal stromal cells from human bone marrow. Cytotherapy, 11:153–162.
27.
DolezelJ, BartosJ, VoglmayrH and GreilhuberJ. (2003). Nuclear DNA content and genome size of trout and human. Cytometry A, 51:127–8; author reply 129.
28.
ZscharnackM, PoeselC, GalleJ and BaderA. (2009). Low oxygen expansion improves subsequent chondrogenesis of ovine bone-marrow-derived mesenchymal stem cells in collagen type I hydrogel. Cells Tissues Organs, 190:81–93.
29.
McGonagleD, BaboolalTG and JonesE. (2017). Native joint-resident mesenchymal stem cells for cartilage repair in osteoarthritis. Nat Rev Rheumatol, 13:719–730.
30.
HuangJI, ZukPA, JonesNF, ZhuM, LorenzHP, HedrickMH and BenhaimP. (2004). Chondrogenic potential of multipotential cells from human adipose tissue. Plast Reconstr Surg, 113:585–594.
31.
RonzièreMC, PerrierE, Mallein-GerinF and FreyriaA-M. (2010). Chondrogenic potential of bone marrow- and adipose tissue-derived adult human mesenchymal stem cells. Biomed Mater Eng, 20:145–158.
32.
AfizahH, YangZ, HuiJHP, OuyangH-W and LeeE-H. (2007). A comparison between the chondrogenic potential of human bone marrow stem cells (BMSCs) and adipose-derived stem cells (ADSCs) taken from the same donors. Tissue Eng, 13:659–666.
33.
WinterA, BreitS, ParschD, BenzK, SteckE, HaunerH, WeberRM, EwerbeckV and RichterW. (2003). Cartilage-like gene expression in differentiated human stem cell spheroids: a comparison of bone marrow-derived and adipose tissue-derived stromal cells. Arthritis Rheum, 48:418–429.
34.
HennigT, LorenzH, ThielA, GoetzkeK, DickhutA, GeigerF and RichterW. (2007). Reduced chondrogenic potential of adipose tissue derived stromal cells correlates with an altered TGFbeta receptor and BMP profile and is overcome by BMP-6. J Cell Physiol, 211:682–691.
35.
SakaguchiY, SekiyaI, YagishitaK and MunetaT. (2005). Comparison of human stem cells derived from various mesenchymal tissues: superiority of synovium as a cell source. Arthritis Rheum, 52:2521–2529.
36.
Hermida-GómezT, Fuentes-BoqueteI, Gimeno-LongasMJ, Muiños-LópezE, Díaz-PradoS, de Toro, JavierFranscisco and BlancoFJ. (2011). Bone marrow cells immunomagnetically selected for CD271+ antigen promote in vitro the repair of articular cartilage defects. Tissue Eng Part A, 17:1169–1179.
37.
KoellingS, KruegelJ, IrmerM, PathJR, SadowskiB, MiroX and MiosgeN. (2009). Migratory chondrogenic progenitor cells from repair tissue during the later stages of human osteoarthritis. Cell Stem Cell, 4:324–335.
38.
CampbellTM, ChurchmanSM, GomezA, McGonagleD, ConaghanPG, PonchelF and JonesE. (2016). Mesenchymal stem cell alterations in bone marrow lesions in patients with hip osteoarthritis. Arthritis Rheum, 68:1648–1659.
39.
Alvarez-ViejoM, Menendez-MenendezY, Blanco-GelazMA, Ferrero-GutierrezA, Fernandez-RodriguezMA, GalaJ and Otero-HernandezJ. (2013). Quantifying mesenchymal stem cells in the mononuclear cell fraction of bone marrow samples obtained for cell therapy. Transplant Proc, 45:434–439.
40.
LiH, GhazanfariR, ZacharakiD, DitzelN, IsernJ, EkblomM, Mendez-FerrerS, KassemM and SchedingS. (2014). Low/negative expression of PDGFR-alpha identifies the candidate primary mesenchymal stromal cells in adult human bone marrow. Stem Cell Reports, 3:965–974.
41.
GarsE, YousrySM, BabuD, KurzerJH, GeorgeTI and GratzingerD. (2017). A replicable CD271+ mesenchymal stromal cell density score: bringing the dysfunctional myelodysplastic syndrome niche to the diagnostic laboratory. Leuk Lymphoma, 58:1730–1732.
42.
Flores-FigueroaE, VarmaS, MontgomeryK, GreenbergPL and GratzingerD. (2012). Distinctive contact between CD34+ hematopoietic progenitors and CXCL12+ CD271+ mesenchymal stromal cells in benign and myelodysplastic bone marrow. Lab Invest, 92:1330–1341.
43.
BusserH, NajarM, RaicevicG, PietersK, Velez PomboR, PhilippartP, MeulemanN, BronD and LagneauxL. (2015). Isolation and characterization of human mesenchymal stromal cell subpopulations: comparison of bone marrow and adipose tissue. Stem Cells Dev, 24:2142–2157.
44.
CuthbertRJ, GiannoudisPV, WangXN, NicholsonL, PawsonD, LubenkoA, TanHB, DickinsonA, McGonagleD and JonesE. (2015). Examining the feasibility of clinical grade CD271+ enrichment of mesenchymal stromal cells for bone regeneration. PLoS One, 10:e0117855.
45.
StubendorffJJ, LammentaustaE, StruglicsA, LindbergL, HeinegårdD and DahlbergLE. (2012). Is cartilage sGAG content related to early changes in cartilage disease? Implications for interpretation of dGEMRIC. Osteoarthritis Cartilage, 20:396–404.
46.
KuiperNJ and SharmaA. (2015). A detailed quantitative outcome measure of glycosaminoglycans in human articular cartilage for cell therapy and tissue engineering strategies. Osteoarthritis Cartilage, 23:2233–2241.
47.
KimIL, MauckRL and BurdickJA. (2011). Hydrogel design for cartilage tissue engineering: a case study with hyaluronic acid. Biomaterials, 32:8771–8782.
48.
VegaSL, KwonMY and BurdickJA. (2017). Recent advances in hydrogels for cartilage tissue engineering. Eur Cell Mater, 33:59–75.
49.
SchneiderU, Schmidt-RohlfingB, GavenisK, MausU, Mueller-RathR and AndereyaS. (2011). A comparative study of 3 different cartilage repair techniques. Knee Surg Sports Traumatol Arthrosc, 19:2145–2152.
50.
SchüttlerKF, SchenkerH, TheisenC, SchoferMD, GetgoodA, RoesslerPP, StruewerJ, RomingerMB and EfeT. (2014). Use of cell-free collagen type I matrix implants for the treatment of small cartilage defects in the knee: clinical and magnetic resonance imaging evaluation. Knee Surg Sports Traumatol Arthrosc, 22:1270–1276.
51.
DexheimerV, FrankS and RichterW. (2012). Proliferation as a requirement for in vitro chondrogenesis of human mesenchymal stem cells. Stem Cells Dev, 21:2160–2169.
52.
NiemeyerP, LauteV, JohnT, BecherC, DiehlP, KolombeT, FayJ, SieboldR, NiksM, FickertS and ZinserW. (2016). The effect of cell dose on the early magnetic resonance morphological outcomes of autologous cell implantation for articular cartilage defects in the knee: a randomized clinical trial. Am J Sports Med, 44:2005–2014.
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