DOI QR코드

DOI QR Code

The effect of serum types on Chondrogenic differentiation of adipose-derived stem cells

  • Cho, Hyeran (Division of Bioengineering, College of Life Sciences and Bioengineering, Incheon National University) ;
  • Lee, Aeri (Division of Bioengineering, College of Life Sciences and Bioengineering, Incheon National University) ;
  • Kim, Kyobum (Division of Bioengineering, College of Life Sciences and Bioengineering, Incheon National University)
  • Received : 2017.11.29
  • Accepted : 2018.02.14
  • Published : 2018.03.01

Abstract

Background: Fetal bovine serum (FBS) is the most essential supplement in culture media for cellular proliferation, metabolism, and differentiation. However, due to a limited supply and subsequently rising prices, a series of studies have investigated a biological feasibility of replaceable serums to substitute FBS. Along with the increasing interests to manufacture stem cell-based cellular products, optimizing the composition of culture media including serums and exogenous growth factors (GFs) is of importance. In this experiment, the effect of bovine serum (BS) and newborn calf serum (NCS) on proliferation and chondrogenic differentiation capacity of human adipose derived stem cells (ADSCs) was evaluated, especially in the chondrogenically supplemented culture condition. Methods: ADSCs were chondrogenically cultured with FBS, BS, and NCS for 14 days. For the acceleration of in vitro chondrogenesis, exogenous insulin-like growth factor and transforming growth $factor-{\beta}3$ were added. Viability and proliferation of ADSCs were evaluated using Live/Dead fluorescence staining and DNA amount, respectively. To investigate a chondrogenic differentiation, a series of assays were performed including a quantification of glycosaminoglycan deposition, alcian blue staining, and RT-PCR analysis for type II collagen, aggrecan and Sox-9 genes. Results: The results demonstrated that proliferation of ADSCs was facilitated in FBS condition as compared with other serum types. For chondrogenic marker gene expression, serum substitutes enhanced Sox-9 expression level on day 14. The deposition of glycosaminoglycan was more facilitated in BS condition regardless of additional chondrogenic GFs. Conclusion: It could be presumably speculated that serum types and exogenous supplements of GFs could also be important parameters to optimize culture media composition, especially in order to maintain the enhanced levels of both proliferation and chondrogenic differentiation of ADSCs during expansion.

Keywords

Acknowledgement

Supported by : National Research Foundation of Korea

References

  1. Lai JH, Rogan H, Kajiyama G, Goodman SB, Smith RL, Maloney W, Yang F. Interaction between osteoarthritic chondrocytes and adipose-derived stem cells is dependent on cell distribution in three-dimension and transforming growth factor-beta3 induction. Tissue Eng A. 2015;21(5-6):992-1002. https://doi.org/10.1089/ten.tea.2014.0244
  2. Waters HA, Geffre CP, Gonzales DA, Grana WA, Szivek JA. Co-culture of adipose derived stem cells and chondrocytes with surface modifying proteins induces enhanced cartilage tissue formation. Journal of investigative surgery : the official journal of the Academy of Surgical Research. 2013;26(3):118-26. https://doi.org/10.3109/08941939.2012.728681
  3. Jo CH, Lee YG, Shin WH, Kim H, Chai JW, Jeong EC, Kim JE, Shim H, Shin JS, Shin IS, Ra JC, Oh S, Yoon KS. Intra-articular injection of mesenchymal stem cells for the treatment of osteoarthritis of the knee: a proof-of-concept clinical trial. Stem Cells. 2014;32(5):1254-66. https://doi.org/10.1002/stem.1634
  4. Kim YS, Koh YG. Injection of mesenchymal stem cells as a supplementary strategy of marrow stimulation improves cartilage regeneration after lateral sliding calcaneal osteotomy for Varus ankle osteoarthritis: clinical and second-look arthroscopic results. Arthroscopy. 2016;32(5):878-89. https://doi.org/10.1016/j.arthro.2016.01.020
  5. Kim YS, Choi YJ, Suh DS, Heo DB, Kim YI, Ryu JS, Koh YG. Mesenchymal stem cell implantation in osteoarthritic knees: is fibrin glue effective as a scaffold? Am J Sports Med. 2015;43(1):176-85. https://doi.org/10.1177/0363546514554190
  6. Qi YY, Du Y, Li WX, Dai XS, Zhao TF, Yan WQ. Cartilage repair using mesenchymal stem cell (MSC) sheet and MSCs-loaded bilayer PLGA scaffold in a rabbit model. Knee Surg Sport Tr A. 2014;22(6):1424-33. https://doi.org/10.1007/s00167-012-2256-3
  7. Naderi N, Combellack EJ, Griffin M, Sedaghati T, Javed M, Findlay MW, Wallace CG, Mosahebi A, Butler PEM, Seifalian AM, Whitaker IS. The regenerative role of adipose-derived stem cells (ADSC) in plastic and reconstructive surgery. Int Wound J. 2017;14(1):112-24. https://doi.org/10.1111/iwj.12569
  8. Lam J, Lu S, Kasper FK, Mikos AG. Strategies for controlled delivery of biologics for cartilage repair. Adv Drug Deliver Rev. 2015;84:123-34. https://doi.org/10.1016/j.addr.2014.06.006
  9. Nukavarapu SP, Dorcemus DL. Osteochondral tissue engineering: current strategies and challenges. Biotechnol Adv. 2013;31(5):706-21. https://doi.org/10.1016/j.biotechadv.2012.11.004
  10. Shen J, Gao QF, Zhang Y, He YH. Autologous platelet-rich plasma promotes proliferation and chondrogenic differentiation of adipose-derived stem cells. Mol Med Rep. 2015;11(2):1298-303. https://doi.org/10.3892/mmr.2014.2875
  11. Burke J, Hunter M, Kolhe R, Isales C, Hamrick M, Fulzele S. Therapeutic potential of mesenchymal stem cell based therapy for osteoarthritis. Clin Transl Med. 2016;5
  12. Ude CC, Sulaiman SB, Ng MH, Chen HC, Ahmad J, Yahaya NM, Saim AB, Idrus RBH. Cartilage regeneration by Chondrogenic induced adult stem cells in osteoarthritic sheep model. PLoS One. 2014;9(6)e98770. https://doi.org/10.1371/journal.pone.0098770
  13. Pak J. Regeneration of human bones in hip osteonecrosis and human cartilage in knee osteoarthritis with autologous adipose-tissue-derived stem cells: a case series. J Med Case Rep. 2011;5:296. https://doi.org/10.1186/1752-1947-5-296
  14. Koh YG, Choi YJ, Kwon SK, Kim YS, Yeo JE. Clinical results and second-look arthroscopic findings after treatment with adipose-derived stem cells for knee osteoarthritis. Knee Surg Sport Tr A. 2015;23(5):1308-16. https://doi.org/10.1007/s00167-013-2807-2
  15. Pak J, Chang JJ, Lee JH, Lee SH. Safety reporting on implantation of autologous adipose tissue-derived stem cells with platelet-rich plasma into human articular joints. Bmc Musculoskel Dis. 2013;14
  16. Singh M, Pierpoint M, Mikos AG, Kasper FK. Chondrogenic differentiation of neonatal human dermal fibroblasts encapsulated in alginate beads with hydrostatic compression under hypoxic conditions in the presence of bone morphogenetic protein-2. J Biomed Mater Res A. 2011;98A(3):412-24. https://doi.org/10.1002/jbm.a.33129
  17. Goldring MB, Tsuchimochi K, Ijiri K. The control of chondrogenesis. J Cell Biochem. 2006;97(1):33-44. https://doi.org/10.1002/jcb.20652
  18. Lund P, Pilgaard L, Duroux M, Fink T, Zachar V. Effect of growth media and serum replacements on the proliferation and differentiation of adiposederived stem cells. Cytotherapy. 2009;11(2):189-97. https://doi.org/10.1080/14653240902736266
  19. Fang CY, Wu CC, Fang CL, Chen WY, Chen CL. Long-term growth comparison studies of FBS and FBS alternatives in six head and neck cell lines. PLoS One. 2017;12(6)
  20. Goedecke A, Wobus M, Krech M, Munch N, Richter K, Holig K, Bornhauser M. Differential effect of platelet-rich plasma and fetal calf serum on bone marrowderived human mesenchymal stromal cells expanded in vitro. J Tissue Eng Regen M. 2011;5(8):648-54. https://doi.org/10.1002/term.359
  21. Mahamoud A, Osman HA, Mansour D, el Harith A. Successful substitution of fetal calf serum by human plasma for bulk cultivation of Leishmania donovani promastigotes. J Med Microbiol. 2013;62:1165-9. https://doi.org/10.1099/jmm.0.052993-0
  22. Murray CF, Leslie KE. Newborn calf vitality: risk factors, characteristics, assessment, resulting outcomes and strategies for improvement. Vet J. 2013;198(2):322-8. https://doi.org/10.1016/j.tvjl.2013.06.007
  23. Hoshiyama Y, Otsuki S, Oda S, Kurokawa Y, Nakajima M, Jotoku T, Tamura R, Okamoto Y, Lotz MK, Neo M. Chondrocyte clusters adjacent to sites of cartilage degeneration have characteristics of progenitor cells. J Orthop Res. 2015;33(4):548-55. https://doi.org/10.1002/jor.22782
  24. Wang B, Chen MZ. Astragaloside IV possesses antiarthritic effect by preventing interleukin 1beta-induced joint inflammation and cartilage damage. Arch Pharm Res. 2014;37(6):793-802. https://doi.org/10.1007/s12272-014-0336-2
  25. Zhu L, Skoultchi AI. Coordinating cell proliferation and differentiation. Curr Opin Genet Dev. 2001;11(1):91-7. https://doi.org/10.1016/S0959-437X(00)00162-3
  26. Rocha PM, Santo VE, Gomes ME, Reis RL, Mano JF. Encapsulation of adiposederived stem cells and transforming growth factor-beta 1 in carrageenanbased hydrogels for cartilage tissue engineering. J Bioact Compat Pol. 2011;26(5):493-507. https://doi.org/10.1177/0883911511420700
  27. Mishra A, Tummala P, King A, Lee B, Kraus M, Tse V, Jacobs CR. Buffered platelet-rich plasma enhances mesenchymal stem cell proliferation and chondrogenic differentiation. Tissue engineering Part C, Methods. 2009;15(3):431-5. https://doi.org/10.1089/ten.tec.2008.0534
  28. Kim K, Lam J, Lu S, Spicer PP, Lueckgen A, Tabata Y, Wong ME, Jansen JA, Mikos AG, Kasper FK. Osteochondral tissue regeneration using a bilayered composite hydrogel with modulating dual growth factor release kinetics in a rabbit model. J Control Release. 2013;168(2):166-78. https://doi.org/10.1016/j.jconrel.2013.03.013
  29. Mahmoudifar N, Doran PM. Chondrogenic differentiation of human adipose-derived stem cells in polyglycolic acid mesh scaffolds under dynamic culture conditions. Biomaterials. 2010;31(14):3858-67. https://doi.org/10.1016/j.biomaterials.2010.01.090
  30. Liao JH, Guo XA, Grande-Allen KJ, Kasper FK, Mikos AG. Bioactive polymer/extracellular matrix scaffolds fabricated with a flow perfusion bioreactor for cartilage tissue engineering. Biomaterials. 2010;31(34):8911-20. https://doi.org/10.1016/j.biomaterials.2010.07.110
  31. Meretoja VV, Dahlin RL, Kasper FK, Mikos AG. Enhanced chondrogenesis in co-cultures with articular chondrocytes and mesenchymal stem cells. Biomaterials. 2012;33(27):6362-9. https://doi.org/10.1016/j.biomaterials.2012.05.042
  32. Dahlin RL, Meretoja VV, Ni MW, Kasper FK, Mikos AG. Chondrogenic phenotype of articular chondrocytes in monoculture and co-culture with mesenchymal stem cells in flow perfusion. Tissue Eng Pt A. 2014;20(21-22):2883-91. https://doi.org/10.1089/ten.tea.2014.0107
  33. Kim K, Dean D, Wallace J, Breithaupt R, Mikos AG, Fisher JP. The influence of stereolithographic scaffold architecture and composition on osteogenic signal expression with rat bone marrow stromal cells. Biomaterials. 2011;32(15):3750-63. https://doi.org/10.1016/j.biomaterials.2011.01.016
  34. Ichiyama S, Funasaka Y, Otsuka Y, Takayama R, Kawana S, Saeki H, Kubo A. Effective treatment by glycolic acid peeling for cutaneous manifestation of familial generalized acanthosis nigricans caused by FGFR3 mutation. Journal of the European Academy of Dermatology and Venereology : JEADV. 2016;30(3):442-5. https://doi.org/10.1111/jdv.13580
  35. Lee DM, Bajracharya P, Lee EJ, Kim JE, Lee HJ, Chun T, Kim J, Cho KH, Chang J, Hong S, Choi I. Effects of gender-specific adult bovine serum on myogenic satellite cell proliferation, differentiation and lipid accumulation. In vitro cellular & developmental biology Animal. 2011;47(7):438-44. https://doi.org/10.1007/s11626-011-9427-2
  36. Kyllonen L, Haimi S, Mannerstrom B, Huhtala H, Rajala KM, Skottman H, Sandor GK, Miettinen S. Effects of different serum conditions on osteogenic differentiation of human adipose stem cells in vitro. Stem Cell Res Ther. 2013;4(1):17. https://doi.org/10.1186/scrt165
  37. Okubo T, Hayashi D, Yaguchi T, Fujita Y, Sakaue M, Suzuki T, Tsukamoto A, Murayama O, Lynch J, Miyazaki Y, Tanaka K, Takizawa T. Differentiation of rat adipose tissue-derived stem cells into neuron-like cells by valproic acid, a histone deacetylase inhibitor. Exp Anim Tokyo. 2016;65(1):45-51. https://doi.org/10.1538/expanim.15-0038
  38. Zhao LL, Ju DP, Gao Q, Zheng XL, Yang GS. Over-expression of Nkx2.5 and/or cardiac alpha-actin inhibit the contraction ability of ADSCs-derived cardiomyocytes. Mol Biol Rep. 2012;39(3):2585-95. https://doi.org/10.1007/s11033-011-1011-z
  39. Park J, Park J, Nahm SS, Choi I, Kim J. Identification of anti-adipogenic proteins in adult bovine serum suppressing 3T3-L1 preadipocyte differentiation. BMB Rep. 2013;46(12):582-7. https://doi.org/10.5483/BMBRep.2013.46.12.082
  40. Lindroos B, Boucher S, Chase L, Kuokkanen H, Huhtala H, Haataja R, Vemuri M, Suuronen R, Miettinen S. Serum-free, xeno-free culture media maintain the proliferation rate and multipotentiality of adipose stem cells in vitro. Cytotherapy. 2009;11(7):958-72. https://doi.org/10.3109/14653240903233081
  41. Ho ST, Tanavde VM, Hui JH, Lee EH. Upregulation of Adipogenesis and Chondrogenesis in MSC serum-free culture. Cell medicine. 2011;2(1):27-41. https://doi.org/10.3727/215517911X575984
  42. Koellensperger E, Bollinger N, Dexheimer V, Gramley F, Germann G, Leimer U. Choosing the right type of serum for different applications of human adipose tissue-derived stem cells: influence on proliferation and differentiation abilities. Cytotherapy. 2014;16(6):789-99. https://doi.org/10.1016/j.jcyt.2014.01.007
  43. Park H, Temenoff JS, Tabata Y, Caplan AI, Mikos AG. Injectable biodegradable hydrogel composites for rabbit marrow mesenchymal stem cell and growth factor delivery for cartilage tissue engineering. Biomaterials. 2007;28(21):3217-27. https://doi.org/10.1016/j.biomaterials.2007.03.030
  44. Park H, Temenoff JS, Tabata Y, Caplan AI, Raphael RM, Jansen JA, Mikos AG. Effect of dual growth factor delivery on chondrogenic differentiation of rabbit marrow mesenchymal stem cells encapsulated in injectable hydrogel composites. J Biomed Mater Res A. 2009;88A(4):889-97. https://doi.org/10.1002/jbm.a.31948
  45. Shainer R, Gaberman E, Levdansky L, Gorodetsky R. Efficient isolation and chondrogenic differentiation of adult mesenchymal stem cells with fibrin microbeads and micronized collagen sponges. Regen Med. 2010;5(2):255-65. https://doi.org/10.2217/rme.09.90
  46. Dexheimer V, Frank S, Richter W. Proliferation as a requirement for in vitro Chondrogenesis of human mesenchymal stem cells. Stem Cells Dev. 2012;21(12):2160-9. https://doi.org/10.1089/scd.2011.0670
  47. Liu Y, Li YQ, Wang HY, Li YJ, Liu GY, Xu X, Wu XB, Jing YG, Yao Y, Wu CT, Jin JD. Effect of serum choice on replicative senescence in mesenchymal stromal cells. Cytotherapy. 2015;17(7):874-84. https://doi.org/10.1016/j.jcyt.2015.02.012
  48. Estes BT, Diekman BO, Guilak F. Monolayer cell expansion conditions affect the chondrogenic potential of adipose-derived stem cells. Biotechnol Bioeng. 2008;99(4):986-95. https://doi.org/10.1002/bit.21662
  49. Ito A, Aoyama T, Iijima H, Nagai M, Yamaguchi S, Tajino J, Zhang X, Akiyama H, Kuroki H. Optimum temperature for extracellular matrix production by articular chondrocytes. Int J Hyperth. 2014;30(2):96-101. https://doi.org/10.3109/02656736.2014.880857

Cited by

  1. Isolation and biological characteristics of multipotent mesenchymal stromal cells derived from chick embryo intestine vol.59, pp.5, 2018, https://doi.org/10.1080/00071668.2018.1490495
  2. A Non-Enzymatic Method to Obtain a Fat Tissue Derivative Highly Enriched in Adipose Stem Cells (ASCs) from Human Lipoaspirates: Preliminary Results vol.19, pp.7, 2018, https://doi.org/10.3390/ijms19072061
  3. In Vitro Effect of Estradiol and Progesterone on Ovine Amniotic Epithelial Cells vol.2019, pp.None, 2019, https://doi.org/10.1155/2019/8034578
  4. Chondrogenic Differentiation of Pluripotent Stem Cells under Controllable Serum-Free Conditions vol.20, pp.11, 2018, https://doi.org/10.3390/ijms20112711
  5. Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources duringin vitroculture vol.28, pp.1, 2018, https://doi.org/10.1017/s0967199419000583
  6. Extracellular matrix-based biomaterials as adipose-derived stem cell delivery vehicles in wound healing: a comparative study between a collagen scaffold and two xenografts vol.11, pp.1, 2020, https://doi.org/10.1186/s13287-020-02021-x
  7. Bioconversion Products of Whey by Lactic Acid Bacteria Exert Anti-Adipogenic Effect vol.41, pp.1, 2021, https://doi.org/10.5851/kosfa.2020.e78
  8. An integrated microfluidic device for stem cell differentiation based on cell-imprinted substrate designed for cartilage regeneration in a rabbit model vol.121, pp.None, 2018, https://doi.org/10.1016/j.msec.2020.111794
  9. ‘Fat chance’: a review of adipose tissue engineering and its role in plastic and reconstructive surgery vol.103, pp.4, 2018, https://doi.org/10.1308/rcsann.2020.7031