Volume 14, Issue 9 (9-2016)                   IJRM 2016, 14(9): 567-576 | Back to browse issues page

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Esmaeli A, Moshrefi M, Shamsara A, Eftekhar-vaghefi S H, Nematollahi-mahani S N. Xeno-free culture condition for human bone marrow and umbilical cord matrix-derived mesenchymal stem/stromal cells using human umbilical cord blood serum. IJRM. 2016; 14 (9) :567-576
URL: http://journals.ssu.ac.ir/ijrmnew/article-1-785-en.html
1- Physiology Research Center, Institute of neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran
2- Neuroscience Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran
3- Department of Anatomy, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran
4- Physiology Research Center, Institute of neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran , nnematollahi@kmu..ac.ir
Abstract:   (968 Views)
Background: Fetal bovine serum (FBS) is widely used in cell culture laboratories,risk of zoonotic infections and allergic side effects create obstacles for its use inclinical trials. Therefore, an alternative supplement with proper inherent growthpromotingactivities is demanded.
Objective: To find FBS substitute, we tested human umbilical cord blood serum(hUCS) for proliferation of human umbilical cord matrix derived mesenchymal stemcells (hUC-MSCs) and human bone marrow-derived mesenchymal cells (hBMMSCs).
Materials and Methods: Umbilical cord blood of healthy neonates, delivered byCaesarian section, was collected and the serum was separated. hUC-MSCs andhBM-MSCs were isolated and characterized by assessment of cell surface antigensby flow cytometry, alkaline phosphatase activity and osteogenic/adipogenicdifferentiation potential. The cells were then cultured in Iscove's ModifiedDulbecco's Medium (IMDM) by conventional methods in three preparations: 1- withhUCS, 2- with FBS, and 3- without serum supplements. Cell proliferation wasmeasured using WST-1 assay, and cell viability was assessed by trypan bluestaining.
Results: The cells cultured in hUCS and FBS exhibited similar morphology andmesenchymal stem cells properties. WST-1 proliferation assay data showed nosignificant difference between the proliferation rate of either cells following hUCSand FBS supplementation. Trypan blue exclusion dye test also revealed nosignificant difference for viability between hUCS and FBS groups. A significantdifference was detected between the proliferation rate of stem cells cultured inserum-supplemented medium compared with serum-free medium.
Conclusion: Our results indicate that human umbilical cord serum can effectivelysupport proliferation of hBM-MSCS and hUC-MSCs in vitro and can be used as anappropriate substitute for FBS, especially in clinical studies.
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Type of Study: Original Article |

1. Walther G, Gekas J, Bertrand OF. Amniotic stem cells for cellular cardiomyoplasty: promises and premises. Catheter Cardiovasc Int 2009; 73: 917-924. [DOI:10.1002/ccd.22016]
2. Shetty P, Cooper K, Viswanathan C. Comparison of proliferative and multilineage differentiation potentials of cord matrix, cord blood, and bone marrow mesenchymal stem cells. Asian J Transfus Sci 2010; 4: 14-24. [DOI:10.4103/0973-6247.59386]
3. Salehinejad P, Alitheen NB, Nematollahi-Mahani SN, Ali AM, Omar AR, Janzamin E, et al. Effect of culture media on expansion properties of human umbilical cord matrix-derived mesenchymal cells. Cytotherapy 2012; 14: 948-953. [DOI:10.3109/14653249.2012.684377]
4. Lindroos B, Aho KL, Kuokkanen H, Raty S, Miettinen S. Differential gene expression in adipose stem cells cultured in allogeneic human serum versus fetal bovine serum. Tissue Eng Part A 2010; 16: 2281-2294. [DOI:10.1089/ten.tea.2009.0621]
5. He S, Nakada D, Morrison SJ. Mechanisms of stem cell self-renewal. Ann Rev cell Dev Biol 2009; 25: 377-406. [DOI:10.1146/annurev.cellbio.042308.113248]
6. Shahdadfar A, Fronsdal K, Haug T, Reinholt FP, Brinchmann JE. In vitro expansion of human mesenchymal stem cells: choice of serum is a determinant of cell proliferation, differentiation, gene expression, and transcriptome stability. Stem Cells 2005; 23: 1357-1366. [DOI:10.1634/stemcells.2005-0094]
7. Latifpour M, Nematollahi-Mahani SN, Deilamy M, Azimzadeh BS, Eftekhar-Vaghefi SH, Nabipour F, et al. Improvement in cardiac function following transplantation of human umbilical cord matrix-derived mesenchymal cells. Cardiology 2011; 120: 9-18. [DOI:10.1159/000332581]
8. Aghaee-Afshar M, Rezazadehkermani M, Asadi A, Malekpour-Afshar R, Shahesmaeili A, Nematollahi-mahani SN. Potential of human umbilical cord matrix and rabbit bone marrow-derived mesenchymal stem cells in repair of surgically incised rabbit external anal sphincter. Dis Colon Rectum 2009; 52: 1753-1761. [DOI:10.1007/DCR.0b013e3181b55112]
9. Lu LL, Song YP, Wei XD, Fang BJ, Li YF. Comparative characterization of mesenchymal stem cells from human umbilical cord tissue and bone marrow. Zhongguo Shi Yan Xue Ye Xue Za Zhi 2008; 16: 140-146.
10. Ryan JM. Effect of different fetal bovine serum concentrations on the replicative life span of cultured chick cells. In Vitro 1979; 15: 895-899. [DOI:10.1007/BF02618046]
11. Fong CY, Biswas A, Tan JH, Gauthaman K, Chan WK, Bongso A. Human Wharton's jelly stem cells have unique transcriptome profiles compared to human embryonic stem cells and other mesenchymal stem cells. Stem Cell Rev Rep 2011; 7: 1-16. [DOI:10.1007/s12015-010-9166-x]
12. Meuleman N, Tondreau T, Delforge A, Dejeneffe M, Massy M, Libertalis M, et al. Human marrow mesenchymal stem cell culture: serum-free medium allows better expansion than classical alpha-MEM medium. Eur J Haematol 2006; 76: 309-316. [DOI:10.1111/j.1600-0609.2005.00611.x]
13. Rahman H, Qasim M, Schultze FC, Oellerich M, A RA. Fetal calf serum heat inactivation and lipopolysaccharide contamination influence the human T lymphoblast proteome and phosphoproteome. Proteome Sci 2011; 9: 71. [DOI:10.1186/1477-5956-9-71]
14. Phadnis SM, Joglekar MV, Venkateshan V, Ghaskadbi SM, Hardikar AA, Bhonde RR. Human umbilical cord blood serum promotes growth, proliferation, as well as differentiation of human bone marrow-derived progenitor cells. In Vitro Cell Dev Biol Anim 2006; 42: 283-286. [DOI:10.1290/0512087]
15. Nakamura T, Inatomi T, Sotozono C, Anq L, Kinoshita S. Transplantation of autologous serum-derived cultivated corneal epithelial equivalents for the treatment of severe ocular surface disease. Ophthalmology 2006; 113: 1765-1772. [DOI:10.1016/j.ophtha.2006.04.030]
16. Heike T, Nakahata T. Ex vivo expansion of hematopoietic stem cells by cytokines. Biochim Biophys Acta 2002; 1592: 313-321. [DOI:10.1016/S0167-4889(02)00324-5]
17. van Hensbergen Y, Schipper LF, Brand A, Slot MC, Welling M, Nauta AJ, et al. Ex vivo culture of human CD34+ cord blood cells with thrombopoietin (TPO) accelerates platelet engraftment in a NOD/SCID mouse model. Exp Hematol 2006; 34: 943-950. [DOI:10.1016/j.exphem.2006.04.009]
18. Lindroos B, Aho KL, Kuokkanen H, Raty S, Huhtala H, Lemp1n R, et al. Differential gene expression in adipose stem cells cultured in allogeneic human serum versus fetal bovine serum. Tissue Eng Part A 2010; 16: 2281-2294. [DOI:10.1089/ten.tea.2009.0621]
19. Nimura A, Muneta T, Koga H, Mochizuki T, Suzuki K, Makino H, et al. Increased proliferation of human synovial mesenchymal stem cells with autologous human serum: comparisons with bone marrow mesenchymal stem cells and with fetal bovine serum. Arthritis Rheum 2008; 58: 501-510. [DOI:10.1002/art.23219]
20. Perez-Ilzarbe M, Diez-Campelo M, Aranda P, Tabera S, Lopez T, del Canizo C, et al. Comparison of ex vivo expansion culture conditions of mesenchymal stem cells for human cell therapy. Transfusion 2009; 49: 1901-1910. [DOI:10.1111/j.1537-2995.2009.02226.x]
21. Rajala K, Lindroos B, Hussein SM, Lappalainen RS, Pekkanen-Mattila M, Inzunza J, et al. A defined and xeno-free culture method enabling the establishment of clinical-grade human embryonic, induced pluripotent and adipose stem cells. PLoS One 2010; 5: e10246. [DOI:10.1371/journal.pone.0010246]
22. Mellado-Damas N, Rodriguez JM, Carmona M, Gonzalez J, Prieto J. Ex-vivo expansion and maturation of CD34-positive hematopoietic progenitors optimization of culture conditions. Leuk Res 1999; 23: 1035-1040. [DOI:10.1016/S0145-2126(99)00126-5]
23. Eslaminejad MB, Rouhi L, Najafi SMA, Baharvand H. [Culture of rat mesenchymal stem cell using peripheral blood-derived plasma as the culture medium supplement]. Sci J Blood Tras Org (Khoon) 2008; 5: 25-37. (In Persian)
24. Salehinejad P, Alitheen NB, Ali AM, Omar AR, Mohit M, Janzamin E, et al. Comparison of different methods for the isolation of mesenchymal stem cells from human umbilical cord Wharton's jelly. In Vitro Cell Dev Biol Anim 2012; 48: 75-83. [DOI:10.1007/s11626-011-9480-x]
25. Motamedi B, Ibrahim TAT, Abdul AB, Allaudin ZA, Moshrefi M, Hajghani M, Nematollahi-Mahani SN. Characteristics of human amniotic epithelial cells and bone marrow mesenchymal stem cells in a parallel study: hanging drop colony formation and doubling time. J Regen Med 2015; 4: 2-10.
26. Nematollahi-mahani SN, Rezazadeh-kermani M, Mehrabani M, Nakhaee N. Cytotoxic effects of Tecurium polium on some established cell lines. Pharm Biol 2007; 45: 295-298. [DOI:10.1080/13880200701214904]
27. Hou T, Xu J, Wu X, Xie Z, Luo F, Zhang Z, et al. Umbilical cord Wharton's Jelly: a new potential cell source of mesenchymal stromal cells for b1 tissue engineering. Tissue Eng Part A 2009; 15: 2325-2334. [DOI:10.1089/ten.tea.2008.0402]
28. Shetty P, Bharucha K, Tanavde V. Human umbilical cord blood serum can replace fetal bovine serum in the culture of mesenchymal stem cells. Cell Biol Int 2007; 31: 293-298. [DOI:10.1016/j.cellbi.2006.11.010]
29. Chakraborty A, Dutta J, Das S, Datta H. Effect of cord blood serum on ex vivo human limbal epithelial cell culture. J Ocul Biol Diseases Inform 2012; 5: 77-82. [DOI:10.1007/s12177-013-9106-5]
30. Shetty P, Viswanathan C. Growth of neural precursor cells using umbilical cord blood serum and a process for the preparation for therapeutic purposes. Google Patents; 2011.
31. Aldahmash A, Haack-Sørensen M, Al-Nbaheen M, Harkness L, Abdallah BM, Kassem M. Human serum is as efficient as fetal bovine serum in supporting proliferation and differentiation of human multipotent stromal (mesenchymal) stem cells in vitro and in vivo. Stem Cell Rev Rep 2011; 7: 860-868. [DOI:10.1007/s12015-011-9274-2]
32. Tateishi K, Ando W, Higuchi C, Hart D, Hashimoto J, Nakata K, et al. Comparison of human serum with fetal bovine serum for expansion and differentiation of human synovial MSC: potential feasibility for clinical applications. Cell Transplant 2008; 17: 549-557. [DOI:10.3727/096368908785096024]
33. Choi J, Chung J-H, Kwon G-Y, Kim K-W, Kim S, Chang H. Effectiveness of autologous serum as an alternative to fetal bovine serum in adipose-derived stem cell engineering. Cell Tissue Bank 2013; 14: 413-422. [DOI:10.1007/s10561-012-9341-1]
34. Gallo R, Gambelli F, Gava B, Sasdelli F, Tell1 V, Masini M, et al. Generation and expansion of multipotent mesenchymal progenitor cells from cultured human pancreatic islets. Cell Death Differentiation 2007; 14: 1860-1871. [DOI:10.1038/sj.cdd.4402199]
35. Paranjape S. Goat serum: an alternative to fetal bovine serum in biomedical research. Indian J Exp Biol 2004; 42: 26-35.
36. Handin RI, Lux SE, Stossel TP. Blood: principles and practice of hematology: Lippincott Williams & Wilkins; 2003.
37. Bieback K, Ha VA-T, Hecker A, Grassl M, Kinzebach S, Solz H, et al. Altered gene expression in human adipose stem cells cultured with fetal bovine serum compared to human supplements. Tissue Eng Part A 2010; 16: 3467-3484. [DOI:10.1089/ten.tea.2009.0727]
38. Van der Valk J, Mellor D, Brands R, Fischer R, Gruber F, Gstraunthaler G, et al. The humane collection of fetal bovine serum and possibilities for serum-free cell and tissue culture. Toxicol Invitro 2004; 18: 1-12. [DOI:10.1016/j.tiv.2003.08.009]
39. Ishiyama M, Tominaga H, Shiga M, Sasamoto K, Ohkura Y, Ueno K. A combined assay of cell viability and in vitro cytotoxicity with a highly water-soluble tetrazolium salt, neutral red and crystal violet. Biol Pharm Bullet 1996; 19: 1518-1520. [DOI:10.1248/bpb.19.1518]
40. Weiss ML, Mitchell KE, Hix JE, Medicetty S, El-Zarkouny SZ, Grieger D, et al. Transplantation of porcine umbilical cord matrix cells into the rat brain. Exp Neurol 2003; 182: 288-299. [DOI:10.1016/S0014-4886(03)00128-6]
41. Matsuda J, Yokota I, Iida M, Murakami T, Naito E, Ito M, et al. Serum leptin concentration in cord blood: relationship to birth weight and gender. J Clin Endocrinol Metab 1997; 82: 1642-1644. [DOI:10.1210/jcem.82.5.4063]
42. Lian R Gao, Yu Chen, Ning K Zhang, Xi L Yang, Hui L Liu, Zhi G Wang, et al. Intracoronary infusion of Wharton's jelly-derived mesenchymal stem cells in acute myocardial infarction: double- blind, randomized controlled trial. BMC Med 2015; 13: 162 [DOI:10.1186/s12916-015-0399-z]

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