Advertisement

Tumor Formation of Adult Stem Cell Transplants in Rodent Arthritic Joints

  • Fanny Chapelin
  • Aman Khurana
  • Mohammad Moneeb
  • Florette K. Gray Hazard
  • Chun Fai Ray Chan
  • Hossein Nejadnik
  • Dita Gratzinger
  • Solomon Messing
  • Jason Erdmann
  • Amitabh Gaur
  • Heike E. Daldrup-Link
Research Article

Abstract

Purpose

While imaging matrix-associated stem cell transplants aimed for cartilage repair in a rodent arthritis model, we noticed that some transplants formed locally destructive tumors. The purpose of this study was to determine the cause for this tumor formation in order to avoid this complication for future transplants.

Procedures

Adipose-derived stem cells (ADSC) isolated from subcutaneous adipose tissue were implanted into 24 osteochondral defects of the distal femur in ten athymic rats and two immunocompetent control rats. All transplants underwent serial magnetic resonance imaging (MRI) up to 6 weeks post-transplantation to monitor joint defect repair. Nine transplants showed an increasing size over time that caused local bone destruction (group 1), while 11 transplants in athymic rats (group 2) and 4 transplants in immunocompetent rats did not. We compared the ADSC implant size and growth rate on MR images, macroscopic features, histopathologic features, surface markers, and karyotypes of these presumed neoplastic transplants with non-neoplastic ADSC transplants.

Results

Implants in group 1 showed a significantly increased two-dimensional area at week 2 (p = 0.0092), 4 (p = 0.003), and 6 (p = 0.0205) compared to week 0, as determined by MRI. Histopathological correlations confirmed neoplastic features in group 1 with significantly increased size, cellularity, mitoses, and cytological atypia compared to group 2. Six transplants in group 1 were identified as malignant chondrosarcomas and three transplants as fibromyxoid sarcomas. Transplants in group 2 and immunocompetent controls exhibited normal cartilage features. Both groups showed a normal ADSC phenotype; however, neoplastic ADSC demonstrated a mixed population of diploid and tetraploid cells without genetic imbalance.

Conclusions

ADSC transplants can form tumors in vivo. Preventive actions to avoid in vivo tumor formations may include karyotyping of culture-expanded ADSC before transplantation. In addition, serial imaging of ADSC transplants in vivo may enable early detection of abnormally proliferating cell transplants.

Key words

Magnetic resonance imaging Stem cell therapy Chondrosarcomas Fibromyxoid sarcomas Osteochondral transplants Malignant tumors in vivo 

Notes

Acknowledgments

The authors would like to acknowledge the imaging support provided by the Stanford Small Animal Imaging Facility (SCI3).

Funding

This work was supported by NIH grant 2R01AR054458 from the National Institute of Arthritis and Musculoskeletal and Skin Diseases (PI Daldrup-Link).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

11307_2018_1218_MOESM1_ESM.pdf (165 kb)
ESM 1 (PDF 164 kb)

References

  1. 1.
    Stem cells entering clinical trials. http://clinicaltrials.gov/ct2/results?term=stem+cells (accessed 05/02/2012)
  2. 2.
    Abudusaimi A, Aihemaitijiang Y, Wang YH, Cui L, Maimaitiming S, Abulikemu M (2011) Adipose-derived stem cells enhance bone regeneration in vascular necrosis of the femoral head in the rabbit. J Int Med Res 39:1852–1860CrossRefPubMedGoogle Scholar
  3. 3.
    Cowan CM, Shi YY, Aalami OO, Chou YF, Mari C, Thomas R, Quarto N, Contag CH, Wu B, Longaker MT (2004) Adipose-derived adult stromal cells heal critical-size mouse calvarial defects. Nat Biotechnol 22:560–567CrossRefPubMedGoogle Scholar
  4. 4.
    Cui L, Liu B, Liu G, Zhang W, Cen L, Sun J, Yin S, Liu W, Cao Y (2007) Repair of cranial bone defects with adipose derived stem cells and coral scaffold in a canine model. Biomaterials 28:5477–5486CrossRefPubMedGoogle Scholar
  5. 5.
    Lin G, Garcia M, Ning H, Banie L, Guo YL, Lue TF, Lin CS (2008) Defining stem and progenitor cells within adipose tissue. Stem Cells Dev 17:1053–1063CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Peng L, Jia Z, Yin X, Zhang X, Liu Y, Chen P, Ma K, Zhou C (2008) Comparative analysis of mesenchymal stem cells from bone marrow, cartilage, and adipose tissue. Stem Cells Dev 17:761–773CrossRefPubMedGoogle Scholar
  7. 7.
    Im GI, Shin YW, Lee KB (2005) Do adipose tissue-derived mesenchymal stem cells have the same osteogenic and chondrogenic potential as bone marrow-derived cells? Osteoarthr Cartil 13:845–853CrossRefPubMedGoogle Scholar
  8. 8.
    Kim HJ, Im GI (2009) Chondrogenic differentiation of adipose tissue-derived mesenchymal stem cells: greater doses of growth factor are necessary. J Orthop Res 27:612–619CrossRefPubMedGoogle Scholar
  9. 9.
    Winter A, Breit S, Parsch D, Benz K, Steck E, Hauner H, Weber RM, Ewerbeck V, Richter W (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 48:418–429CrossRefPubMedGoogle Scholar
  10. 10.
    Chun HJ, Kim YS, Kim BK et al (2011) Transplantation of human adipose-derived stem cells in a rabbit model of traumatic degeneration of lumbar discs. World Neurosurg 78:364–371CrossRefPubMedGoogle Scholar
  11. 11.
    Galle J, Bader A, Hepp P, Grill W, Fuchs B, Kas JA, Krinner A, MarquaB B, Muller K, Schiller J, Schulz RM, von Buttlar M, von der Burg E, Zscharnack M, Loffler M (2010) Mesenchymal stem cells in cartilage repair: state of the art and methods to monitor cell growth, differentiation and cartilage regeneration. Curr Med Chem 17:2274–2291CrossRefPubMedGoogle Scholar
  12. 12.
    Gao J, Yao JQ, Caplan AI (2007) Stem cells for tissue engineering of articular cartilage. Proc Inst Mech Eng H 221:441–450CrossRefPubMedGoogle Scholar
  13. 13.
    Okita K, Ichisaka T, Yamanaka S (2007) Generation of germline-competent induced pluripotent stem cells. Nature 448:313–317CrossRefPubMedGoogle Scholar
  14. 14.
    Peng X, Liu T, Wang Y, Yan Q, Jin H, Li L, Qian Q, Wu M (2012) Wnt/beta-catenin signaling in embryonic stem cell converted tumor cells. J Transl Med 10:196CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Shih CC, Forman SJ, Chu P, Slovak M (2007) Human embryonic stem cells are prone to generate primitive, undifferentiated tumors in engrafted human fetal tissues in severe combined immunodeficient mice. Stem Cells Dev 16:893–902CrossRefPubMedGoogle Scholar
  16. 16.
    Spaeth EL, Dembinski JL, Sasser AK, Watson K, Klopp A, Hall B, Andreeff M, Marini F (2009) Mesenchymal stem cell transition to tumor-associated fibroblasts contributes to fibrovascular network expansion and tumor progression. PLoS One 4:e4992CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Martin-Padura I, Gregato G, Marighetti P, Mancuso P, Calleri A, Corsini C, Pruneri G, Manzotti M, Lohsiriwat V, Rietjens M, Petit JY, Bertolini F (2012) The white adipose tissue used in lipotransfer procedures is a rich reservoir of CD34+ progenitors able to promote cancer progression. Cancer Res 72:325–334CrossRefPubMedGoogle Scholar
  18. 18.
    Garcia S, Bernad A, Martin MC et al (2010) Pitfalls in spontaneous in vitro transformation of human mesenchymal stem cells. Exp Cell Res 316:1648–1650CrossRefPubMedGoogle Scholar
  19. 19.
    Ning H, Lin G, Lue TF, Lin CS (2006) Neuron-like differentiation of adipose tissue-derived stromal cells and vascular smooth muscle cells. Differentiation 74:510–518CrossRefPubMedGoogle Scholar
  20. 20.
    Jo VY, Fletcher CD (2014) WHO classification of soft tissue tumours: an update based on the 2013 (4th) edition. Pathology 46:95–104CrossRefPubMedGoogle Scholar
  21. 21.
    Zuk PA, Zhu M, Ashjian P, de Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13:4279–4295CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    de Lucca EJ, Dhaliwal MK, Furlong CL, Pathak S (1990) A high-resolution G-banding idiogram of Rattus norvegicus chromosomes. Cytobios 62:153–160PubMedGoogle Scholar
  23. 23.
    Khurana A, Nejadnik H, Chapelin F, Lenkov O, Gawande R, Lee S, Gupta SN, Aflakian N, Derugin N, Messing S, Lin G, Lue TF, Pisani L, Daldrup-Link HE (2013) Ferumoxytol: a new, clinically applicable label for stem-cell tracking in arthritic joints with MRI. Nanomedicine (Lond) 8:1969–1983CrossRefGoogle Scholar
  24. 24.
    Strem BM, Hicok KC, Zhu M, Wulur I, Alfonso Z, Schreiber RE, Fraser JK, Hedrick MH (2005) Multipotential differentiation of adipose tissue-derived stem cells. Keio J Med 54:132–141CrossRefPubMedGoogle Scholar
  25. 25.
    Ning H, Liu G, Lin G, Garcia M, Li LC, Lue TF, Lin CS (2009) Identification of an aberrant cell line among human adipose tissue-derived stem cell isolates. Differentiation 77:172–180CrossRefPubMedGoogle Scholar
  26. 26.
    Furlani D, Li W, Pittermann E, Klopsch C, Wang L, Knopp A, Jungebluth P, Thedinga E, Havenstein C, Westien I, Ugurlucan M, Li RK, Ma N, Steinhoff G (2009) A transformed cell population derived from cultured mesenchymal stem cells has no functional effect after transplantation into the injured heart. Cell Transplant 18:319–331CrossRefPubMedGoogle Scholar
  27. 27.
    Miura M, Miura Y, Padilla-Nash HM, Molinolo AA, Fu B, Patel V, Seo BM, Sonoyama W, Zheng JJ, Baker CC, Chen W, Ried T, Shi S (2006) Accumulated chromosomal instability in murine bone marrow mesenchymal stem cells leads to malignant transformation. Stem Cells 24:1095–1103CrossRefPubMedGoogle Scholar
  28. 28.
    Tolar J, Nauta AJ, Osborn MJ, Panoskaltsis Mortari A, McElmurry RT, Bell S, Xia L, Zhou N, Riddle M, Schroeder TM, Westendorf JJ, McIvor RS, Hogendoorn PCW, Szuhai K, Oseth LA, Hirsch B, Yant SR, Kay MA, Peister A, Prockop DJ, Fibbe WE, Blazar BR (2007) Sarcoma derived from cultured mesenchymal stem cells. Stem Cells 25:371–379CrossRefPubMedGoogle Scholar
  29. 29.
    Zhou YF, Bosch-Marce M, Okuyama H, Krishnamachary B, Kimura H, Zhang L, Huso DL, Semenza GL (2006) Spontaneous transformation of cultured mouse bone marrow-derived stromal cells. Cancer Res 66:10849–10854CrossRefPubMedGoogle Scholar
  30. 30.
    Rosland GV, Svendsen A, Torsvik A, Sobala E, McCormack E, Immervoll H, Mysliwietz J, Tonn JC, Goldbrunner R, Lonning PE, Bjerkvig R, Schichor C (2009) Long-term cultures of bone marrow-derived human mesenchymal stem cells frequently undergo spontaneous malignant transformation. Cancer Res 69:5331–5339CrossRefPubMedGoogle Scholar
  31. 31.
    Jeong JO, Han JW, Kim JM, Cho HJ, Park C, Lee N, Kim DW, Yoon YS (2011) Malignant tumor formation after transplantation of short-term cultured bone marrow mesenchymal stem cells in experimental myocardial infarction and diabetic neuropathy. Circ Res 108:1340–1347CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Tapp H, Hanley EN Jr, Patt JC, Gruber HE (2009) Adipose-derived stem cells: characterization and current application in orthopaedic tissue repair. Exp Biol Med (Maywood) 234:1–9CrossRefGoogle Scholar
  33. 33.
    Suga H, Shigeura T, Matsumoto D, Inoue K, Kato H, Aoi N, Murase S, Sato K, Gonda K, Koshima I, Yoshimura K (2007) Rapid expansion of human adipose-derived stromal cells preserving multipotency. Cytotherapy 9:738–745CrossRefPubMedGoogle Scholar
  34. 34.
    Bernardo ME, Zaffaroni N, Novara F, Cometa AM, Avanzini MA, Moretta A, Montagna D, Maccario R, Villa R, Daidone MG, Zuffardi O, Locatelli F (2007) Human bone marrow-derived mesenchymal stem cells do not undergo transformation after long-term in vitro culture and do not exhibit telomere maintenance mechanisms. Cancer Res 67:9142–9149CrossRefPubMedGoogle Scholar
  35. 35.
    Lindroos B, Boucher S, Chase L, Kuokkanen H, Huhtala H, Haataja R, Vemuri M, Suuronen R, Miettinen S (2009) Serum-free, xeno-free culture media maintain the proliferation rate and multipotentiality of adipose stem cells in vitro. Cytotherapy 11:958–972CrossRefPubMedGoogle Scholar
  36. 36.
    Vassaux G, Negrel R, Ailhaud G, Gaillard D (1994) Proliferation and differentiation of rat adipose precursor cells in chemically defined medium: differential action of anti-adipogenic agents. J Cell Physiol 161:249–256CrossRefPubMedGoogle Scholar
  37. 37.
    Lindroos B, Aho KL, Kuokkanen H, Räty S, Huhtala H, Lemponen R, Yli-Harja O, Suuronen R, Miettinen S (2010) Differential gene expression in adipose stem cells cultured in allogeneic human serum versus fetal bovine serum. Tissue Eng Part A 16:2281–2294CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Lund P, Pilgaard L, Duroux M, Fink T, Zachar V (2009) Effect of growth media and serum replacements on the proliferation and differentiation of adipose-derived stem cells. Cytotherapy 11:189–197CrossRefPubMedGoogle Scholar
  39. 39.
    Yoshimura K, Sato K, Aoi N, Kurita M, Hirohi T, Harii K (2008) Cell-assisted lipotransfer for cosmetic breast augmentation: supportive use of adipose-derived stem/stromal cells. Aesthet Plast Surg 32:48–55; discussion 56-47 Google Scholar
  40. 40.
    Alvarez PD, Garcia-Arranz M, Georgiev-Hristov T, Garcia-Olmo D (2008) A new bronchoscopic treatment of tracheomediastinal fistula using autologous adipose-derived stem cells. Thorax 63:374–376CrossRefPubMedGoogle Scholar
  41. 41.
    Lendeckel S, Jodicke A, Christophis P et al (2004) Autologous stem cells (adipose) and fibrin glue used to treat widespread traumatic calvarial defects: case report. J Craniomaxillofac Surg 32:370–373CrossRefPubMedGoogle Scholar
  42. 42.
    Beitzel K, McCarthy MB, Cote MP et al (2012) Rapid isolation of human stem cells (connective progenitor cells) from the distal femur during arthroscopic knee surgery. Arthroscopy 28:74–84CrossRefPubMedGoogle Scholar
  43. 43.
    Tetreault P, Ouellette HA (2007) Healing of a clavicle fracture nonunion with bone marrow injection. J Shoulder Elb Surg 16:e23–e24CrossRefGoogle Scholar
  44. 44.
    Fodor PB, Paulseth SG (2016) Adipose derived stromal cell (ADSC) injections for pain management of osteoarthritis in the human knee joint. Aesthet Surg J 36:229–236CrossRefPubMedGoogle Scholar
  45. 45.
    Ra JC, Jeong EC, Kang SK, Lee SJ, Choi KH (2017) A prospective, nonrandomized, no placebo-controlled, phase I/II clinical trial assessing the safety and efficacy of intramuscular injection of autologous adipose tissue-derived mesenchymal stem cells in patients with severe Buerger’s disease. Cell Med 9:87–102CrossRefPubMedGoogle Scholar
  46. 46.
    Gupta PK, Krishna M, Chullikana A, Desai S, Murugesan R, Dutta S, Sarkar U, Raju R, Dhar A, Parakh R, Jeyaseelan L, Viswanathan P, Vellotare PK, Seetharam RN, Thej C, Rengasamy M, Balasubramanian S, Majumdar AS (2017) Administration of adult human bone marrow-derived, cultured, pooled, allogeneic mesenchymal stromal cells in critical limb ischemia due to Buerger’s disease: phase II study report suggests clinical efficacy. Stem Cells Transl Med 6:689–699CrossRefPubMedGoogle Scholar
  47. 47.
    Wislet-Gendebien S, Poulet C, Neirinckx V, Hennuy B, Swingland JT, Laudet E, Sommer L, Shakova O, Bours V, Rogister B (2012) In vivo tumorigenesis was observed after injection of in vitro expanded neural crest stem cells isolated from adult bone marrow. PLoS One 7:e46425CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Li H, Fan X, Kovi RC, Jo Y, Moquin B, Konz R, Stoicov C, Kurt-Jones E, Grossman SR, Lyle S, Rogers AB, Montrose M, Houghton J (2007) Spontaneous expression of embryonic factors and p53 point mutations in aged mesenchymal stem cells: a model of age-related tumorigenesis in mice. Cancer Res 67:10889–10898CrossRefPubMedGoogle Scholar
  49. 49.
    Wang MY, Nestvold J, Rekdal O, Kvalheim G, Fodstad O (2017) A novel rat fibrosarcoma cell line from transformed bone marrow-derived mesenchymal stem cells with maintained in vitro and in vivo stemness properties. Exp Cell Res 352:218–224CrossRefPubMedGoogle Scholar
  50. 50.
    Tarte K, Gaillard J, Lataillade JJ, Fouillard L, Becker M, Mossafa H, Tchirkov A, Rouard H, Henry C, Splingard M, Dulong J, Monnier D, Gourmelon P, Gorin NC, Sensebe L, on behalf of Societe Francaise de Greffe de Moelle et Therapie Cellulaire (2010) Clinical-grade production of human mesenchymal stromal cells: occurrence of aneuploidy without transformation. Blood 115:1549–1553CrossRefPubMedGoogle Scholar
  51. 51.
    Hahn WC, Weinberg RA (2002) Modelling the molecular circuitry of cancer. Nat Rev Cancer 2:331–341CrossRefPubMedGoogle Scholar
  52. 52.
    Boregowda SV, Krishnappa V, Chambers JW, Lograsso PV, Lai WT, Ortiz LA, Phinney DG (2012) Atmospheric oxygen inhibits growth and differentiation of marrow-derived mouse mesenchymal stem cells via a p53-dependent mechanism: implications for long-term culture expansion. Stem Cells 30:975–987CrossRefPubMedPubMedCentralGoogle Scholar
  53. 53.
    Prockop DJ, Keating A (2012) Relearning the lessons of genomic stability of human cells during expansion in culture: implications for clinical research. Stem Cells 30:1051–1052CrossRefPubMedGoogle Scholar
  54. 54.
    Rubin H (2001) Multistage carcinogenesis in cell culture. Dev Biologicals 106:61–66; discussion 67, 143–160Google Scholar
  55. 55.
    Amariglio N, Hirshberg A, Scheithauer BW, Cohen Y, Loewenthal R, Trakhtenbrot L, Paz N, Koren-Michowitz M, Waldman D, Leider-Trejo L, Toren A, Constantini S, Rechavi G (2009) Donor-derived brain tumor following neural stem cell transplantation in an ataxia telangiectasia patient. PLoS Med 6:e1000029CrossRefPubMedPubMedCentralGoogle Scholar
  56. 56.
    Danylesko I, Shimoni A (2018) Second malignancies after hematopoietic stem cell transplantation. Curr Treat Options in Oncol 19:9CrossRefGoogle Scholar
  57. 57.
    Rizzo JD, Curtis RE, Socie G et al (2009) Solid cancers after allogeneic hematopoietic cell transplantation. Blood 113:1175–1183CrossRefPubMedPubMedCentralGoogle Scholar
  58. 58.
    Curtis RE, Metayer C, Rizzo JD, Socié G, Sobocinski KA, Flowers ME, Travis WD, Travis LB, Horowitz MM, Deeg HJ (2005) Impact of chronic GVHD therapy on the development of squamous-cell cancers after hematopoietic stem-cell transplantation: an international case-control study. Blood 105:3802–3811CrossRefPubMedPubMedCentralGoogle Scholar
  59. 59.
    Garcia-Olmo D, Herreros D, De-La-Quintana P, et al. (2010) Adipose-derived stem cells in Crohn’s rectovaginal fistula. Case Report Med 2010:961758, 1, 3Google Scholar
  60. 60.
    Lee HC, An SG, Lee HW, Park JS, Cha KS, Hong TJ, Park JH, Lee SY, Kim SP, Kim YD, Chung SW, Bae YC, Shin YB, Kim JI, Jung JS (2012) Safety and effect of adipose tissue-derived stem cell implantation in patients with critical limb ischemia. Circ J 76:1750–1760CrossRefPubMedGoogle Scholar
  61. 61.
    Mesimaki K, Lindroos B, Tornwall J et al (2009) Novel maxillary reconstruction with ectopic bone formation by GMP adipose stem cells. Int J Oral Maxillofac Surg 38:201–209CrossRefPubMedGoogle Scholar
  62. 62.
    Scuderi N, Ceccarelli S, Onesti MG, et al. (2012) Human adipose-derived stem cells for cell-based therapies in the treatment of systemic sclerosis. Cell TransplantGoogle Scholar

Copyright information

© World Molecular Imaging Society 2018

Authors and Affiliations

  • Fanny Chapelin
    • 1
  • Aman Khurana
    • 1
  • Mohammad Moneeb
    • 1
  • Florette K. Gray Hazard
    • 2
  • Chun Fai Ray Chan
    • 3
  • Hossein Nejadnik
    • 1
  • Dita Gratzinger
    • 2
  • Solomon Messing
    • 4
  • Jason Erdmann
    • 5
  • Amitabh Gaur
    • 3
    • 6
  • Heike E. Daldrup-Link
    • 1
  1. 1.Department of Radiology, Molecular Imaging Program at Stanford (MIPS)Stanford UniversityStanfordUSA
  2. 2.Department of PathologyStanford UniversityStanfordUSA
  3. 3.BD biosciences, Custom Technology TeamLa JollaUSA
  4. 4.Department of Communication and StatisticsStanfordUSA
  5. 5.Department of CytogeneticsStanford UniversityStanfordUSA
  6. 6.Innovative Assay SolutionsSan DiegoUSA

Personalised recommendations