Human Umbilical Cord-derived Mesenchymal Stem Cells Do Not Undergo Malignant Transformation During Long-term Culturing in Serum-free Medium

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2014 Jun 3

PMID 24887492

Citations 32

Authors

Gecai Chen

Aihuan Yue

Zhongbao Ruan

Yigang Yin

Ruzhu Wang

Yin Ren

Li Zhu

Affiliations

Soon will be listed here.

Abstract

Background: Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are in the foreground as a preferable application for treating diseases. However, the safety of hUC-MSCs after long-term culturing in vitro in serum-free medium remains unclear.

Methods: hUC-MSCs were separated by adherent tissue culture. hUC-MSCs were cultured in serum-free MesenCult-XF medium and FBS-bases DMEM complete medium. At the 1st, 3rd, 5th, 8th, 10th, and 15th passage, the differentiation of MSCs into osteogenic, chondrogenic, and adipogenic cells was detected, and MTT, surface antigens were measured. Tumorigenicity was analyzed at the 15th passage. Conventional karyotyping was performed at passage 0, 8, and 15. The telomerase activity of hUC-MSCs at passage 1-15 was analyzed.

Results: Flow cytometry analysis showed that very high expression was detected for CD105, CD73, and CD90 and very low expression for CD45, CD34, CD14, CD79a, and HLA-DR. MSCs could differentiate into osteocytes, chondrocytes, and adipocytes in vitro. There was no obvious chromosome elimination, displacement, or chromosomal imbalance as determined from the guidelines of the International System for Human Cytogenetic Nomenclature. Telomerase activity was down-regulated significantly when the culture time was prolonged. Further, no tumors formed in rats injected with hUC-MSCs (P15) cultured in serum-free and in serum-containing conditions.

Conclusion: Our data showed that hUC-MSCs met the International Society for Cellular Therapy standards for conditions of long-term in vitro culturing at P15. Since hUC-MSCs can be safely expanded in vitro and are not susceptible to malignant transformation in serum-free medium, these cells are suitable for cell therapy.

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References

Wang L, Ott L, Seshareddy K, Weiss M, Detamore M . Musculoskeletal tissue engineering with human umbilical cord mesenchymal stromal cells. Regen Med. 2010; 6(1):95-109. PMC: 3057462. DOI: 10.2217/rme.10.98. View

Sarugaser R, Lickorish D, Baksh D, Hosseini M, Davies J . Human umbilical cord perivascular (HUCPV) cells: a source of mesenchymal progenitors. Stem Cells. 2005; 23(2):220-9. DOI: 10.1634/stemcells.2004-0166. View

Forostyak S, Jendelova P, Sykova E . The role of mesenchymal stromal cells in spinal cord injury, regenerative medicine and possible clinical applications. Biochimie. 2013; 95(12):2257-70. DOI: 10.1016/j.biochi.2013.08.004. View

Sekulovic S, Gylfadottir V, Vulto I, Gasparetto M, Even Y, Brookes C . Prolonged self-renewal activity unmasks telomerase control of telomere homeostasis and function of mouse hematopoietic stem cells. Blood. 2011; 118(7):1766-73. PMC: 3158711. DOI: 10.1182/blood-2010-11-319632. View

Redaelli S, Bentivegna A, Foudah D, Miloso M, Redondo J, Riva G . From cytogenomic to epigenomic profiles: monitoring the biologic behavior of in vitro cultured human bone marrow mesenchymal stem cells. Stem Cell Res Ther. 2012; 3(6):47. PMC: 3580477. DOI: 10.1186/scrt138. View

Zhou Y, Bosch-Marce M, Okuyama H, Krishnamachary B, Kimura H, Zhang L . Spontaneous transformation of cultured mouse bone marrow-derived stromal cells. Cancer Res. 2006; 66(22):10849-54. DOI: 10.1158/0008-5472.CAN-06-2146. View

Akiyama K, You Y, Yamaza T, Chen C, Tang L, Jin Y . Characterization of bone marrow derived mesenchymal stem cells in suspension. Stem Cell Res Ther. 2012; 3(5):40. PMC: 3580431. DOI: 10.1186/scrt131. View

Torsvik A, Rosland G, Svendsen A, Molven A, Immervoll H, McCormack E . Spontaneous malignant transformation of human mesenchymal stem cells reflects cross-contamination: putting the research field on track - letter. Cancer Res. 2010; 70(15):6393-6. DOI: 10.1158/0008-5472.CAN-10-1305. View

Bernardo M, Zaffaroni N, Novara F, Cometa A, Avanzini M, Moretta A . 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. 2007; 67(19):9142-9. DOI: 10.1158/0008-5472.CAN-06-4690. View

10.

Tang Q, Chen Q, Lai X, Liu S, Chen Y, Zheng Z . Malignant transformation potentials of human umbilical cord mesenchymal stem cells both spontaneously and via 3-methycholanthrene induction. PLoS One. 2013; 8(12):e81844. PMC: 3858282. DOI: 10.1371/journal.pone.0081844. View

11.

Tolar J, Nauta A, Osborn M, Panoskaltsis Mortari A, McElmurry R, Bell S . Sarcoma derived from cultured mesenchymal stem cells. Stem Cells. 2006; 25(2):371-9. DOI: 10.1634/stemcells.2005-0620. View

12.

Al-Nbaheen M, Vishnubalaji R, Ali D, Bouslimi A, Al-Jassir F, Megges M . Human stromal (mesenchymal) stem cells from bone marrow, adipose tissue and skin exhibit differences in molecular phenotype and differentiation potential. Stem Cell Rev Rep. 2012; 9(1):32-43. PMC: 3563956. DOI: 10.1007/s12015-012-9365-8. View

13.

Torsvik A, Rosland G, Bjerkvig R . Comment to: "Spontaneous transformation of adult mesenchymal stem cells from cynomolgus macaques in vitro" by Z. Ren et al. Exp. Cell Res. 317 (2011) 2950-2957: spontaneous transformation of mesenchymal stem cells in culture: facts or fiction?. Exp Cell Res. 2012; 318(5):441-3. DOI: 10.1016/j.yexcr.2011.12.009. View

14.

Aguilar S, Nye E, Chan J, Loebinger M, Spencer-Dene B, Fisk N . Murine but not human mesenchymal stem cells generate osteosarcoma-like lesions in the lung. Stem Cells. 2007; 25(6):1586-94. DOI: 10.1634/stemcells.2006-0762. View

15.

Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D . Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006; 8(4):315-7. DOI: 10.1080/14653240600855905. View

16.

Sensebe L . Clinical grade production of mesenchymal stem cells. Biomed Mater Eng. 2008; 18(1 Suppl):S3-10. View

17.

Herberts C, Kwa M, Hermsen H . Risk factors in the development of stem cell therapy. J Transl Med. 2011; 9:29. PMC: 3070641. DOI: 10.1186/1479-5876-9-29. View

18.

Ahrari I, Purhabibi Zarandi N, Khosravi Maharlooei M, Monabati A, Attari A, Ahrari S . Adipose Tissue Derived Multipotent Mesenchymal Stromal Cells Can Be Isolated Using Serum-free Media. Iran Red Crescent Med J. 2013; 15(4):324-9. PMC: 3785908. DOI: 10.5812/ircmj.4506. View

19.

Purandare B, Teklemariam T, Zhao L, Hantash B . Temporal HLA profiling and immunomodulatory effects of human adult bone marrow- and adipose-derived mesenchymal stem cells. Regen Med. 2013; 9(1):67-79. DOI: 10.2217/rme.13.82. View

20.

Patel D, Shah J, Srivastava A . Therapeutic potential of mesenchymal stem cells in regenerative medicine. Stem Cells Int. 2013; 2013:496218. PMC: 3615627. DOI: 10.1155/2013/496218. View