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THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR

Yıl 2019, , 179 - 190, 31.07.2019
https://doi.org/10.18036/estubtdc.598774

Öz



ABSTRACT



 



Packed-bed bioreactors provide
larger surface area to volume ratio compared to the static culture on flasks.
Therefore, these systems offer ideal production environment for large-scale
culture of mesenchymal stem cells (MSCs), but the effect of fluid dynamics on
the cell-behavior of MSCs is not fully elucidated. In this study, packed-bed
perfusion reactor loaded with different size of polymethyl methacrylate
carriers was used to apply different rates of shear stress and drug forces at
constant flow rate. The cell viability, cell-expansion, apoptosis and protein
secretion levels were analyzed for both unmodified and Vascular Endothelial
Growth Factor-positive (VEGF+) MSCs. The superficial stress was
estimated to between 0.21-0.25 N/m2. The results showed that the
shear stress reduced the VEGF secretion, and Caspase-3 was activated at high
drag force, which cause the reduction of the cell numbers in the bioreactor.
The reduction of cytoskeletal actin structures seemed to play the central role
in this adverse effect of the non-planar shear stress.  The expression reduction of VEGF might also
have critical impacts on the tissue engineering applications, in which the
formation of vascular construct is essential.

Kaynakça

  • [1] Zhang ZY, Teoh SH, Teo EY, Khoon Chong MS, Shin CW, Tien FT, Choolani MA, Chan JK. A comparison of bioreactors for culture of fetal mesenchymal stem cells for bone tissue engineering. Biomaterials 2010; 31: 8684-8695.
  • [2] Bancroft GN, Sikavitsas VI, Mikos AG. Design of a flow perfusion bioreactor system for bone tissue-engineering applications. Tissue Eng 2003; 9: 549-554.
  • [3] Kavlock KD, Goldstein AS. Effect of pulse frequency on the osteogenic differentiation of mesenchymal stem cells in a pulsatile perfusion bioreactor. J Biomech Eng 2011; 133: 091005.
  • [4] Sikavitsas VI, Bancroft GN, Holtorf HL, Jansen JA, Mikos AG. Mineralized matrix deposition by marrow stromal osteoblasts in 3D perfusion culture increases with increasing fluid shear forces. Proc Natl Acad Sci USA 2003; 100: 14683–14688.
  • [5] Osiecki MJ, McElwain SDL, Lott WB. Modelling mesenchymal stromal cell growth in a packed bed bioreactor with a gas permeable wall. PLoS One 2018; 13: e0202079.
  • [6] Zhao F, Chella R, Ma T. Effects of shear stress on 3-D human mesenchymal stem cell construct development in a perfusion bioreactor system: Experiments and hydrodynamic modeling. Biotechnol Bioeng 2007; 96: 584-595.
  • [7] Le Blanc K, Davies LC. MSCs-cells with many sides. Cytotherapy 2018; 20: 273-278.
  • [8] Guo T, Yu L, Lim CG, Goodley AS, Xiao X, Placone JK, Ferlin KM, Nguyen BN, Hsieh AH, Fisher JP. Effect of Dynamic Culture and Periodic Compression on Human Mesenchymal Stem Cell Proliferation and Chondrogenesis. Ann Biomed Eng 2016; 44: 2103-2113.
  • [9] Caplan AI. Adult mesenchymal stem cells for tissue engineering versus regenerative medicine. J Cell Physiol 2007; 213: 341–347.
  • [10] Knippenberg M, Helder MN, Doulabi BZ, Semeins CM, Wuisman PI, Klein-Nulend J. Adipose tissue-derived mesenchymal stem cells acquire bone cell-like responsiveness to fluid shear stress on osteogenic stimulation. Tissue Eng 2005; 11: 1780-1788.
  • [11] Huang Y, Jia X, Bai K, Gong X, Fan Y. Effect of fluid shear stress on cardiomyogenic differentiation of rat bone marrow mesenchymal stem cells. Arch Med Res 2010; 41: 497-505.
  • [12] Ghadirian E., Arastoopour H. CFD simulation of a fluidized bed using the EMMS approach for the gas-solid drag force. Powder Technology 2016; 288: 35-44.
  • [13] Wiklund K, Zhang H, Stangner T, Singh B, Bullitt E, Andersson M. A drag force interpolation model for capsule-shaped cells in fluid flows near a surface. Microbiology. 2018;164(4):483-494.
  • [14] Turac G, Duruksu G, Karaoz E. The Effect of Recombinant Tyrosine Hydroxylase Expression on the Neurogenic Differentiation Potency of Mesenchymal Stem Cells. Neurospine 2018; 15: 42-53.
  • [15] Adas G, Percem A, Adas M, Kemik O, Arikan S, Ustek D, Cakiris A, Abaci N, Kemik AS, Kamali G, et al. VEGF-A and FGF gene therapy accelerate healing of ischemic colonic anastomoses (experimental study). Int J Surg 2011; 9: 467-471.
  • [16] Geankoplis CJ. Transport Processes and Unit Operations. 3rd ed. Englewood Cliffs, NJ, USA: Prentice Hall Press, 1993.
  • [17] Bueno EM, Ruberti JW. Optimizing Collagen Transport Through Track-Etched Nanopores.J Memb Sci 2008; 321: 250-263.
  • [18] Matsuura T. Bioreactors for 3-dimensional high-density culture of human cells. Hum Cell 2006; 19: 11-16.
  • [19] Mitra D, Whitehead J, Yasui OW, Leach JK. Bioreactor culture duration of engineered constructs influences bone formation by mesenchymal stem cells. Biomaterials 2017; 146: 29-39.
  • [20] Li YJ, Batra NN, You LD, Meier SC, Coe IA, Yellowley CE, Jacobs CR. Oscillatory fluid flow affects human marrow stromal cell proliferation and differentiation. J Orthop Res 2004; 22: 1283–1289.
  • [21] Hosseinkhani H, Inatsugu Y, Hiraoka Y, Inoue S, Tabata Y. Perfusion culture enhances osteogenic differentiation of rat mesenchymal stem cells in collagen sponge reinforced with poly(glycolic acid) fiber. Tissue Eng 2005; 11: 1476–1488.
  • [22] Yourek G, McCormick SM, Mao JJ, Reilly GC. Shear stress induces osteogenic differentiation of human mesenchymal stem cells. Regen Med 2010; 5: 713-724.
  • [23] Kim SH, Ahn K, Park JY. Responses of human adipose-derived stem cells to interstitial level of extremely low shear flows regarding differentiation, morphology, and proliferation. Lab Chip 2017; 17(12): 2115-2124.
  • [24] Dong JD, Gu YQ, Li CM, Wang CR, Feng ZG, Qiu RX, Chen B, Li JX, Zhang SW, Wang ZG, et al. Response of mesenchymal stem cells to shear stress in tissue-engineered vascular grafts. Acta Pharmacol Sin 2009; 30: 530-536.
  • [25] Bhaskar B, Owen R, Bahmaee H, Rao PS, Reilly GC. Design and Assessment of a Dynamic Perfusion Bioreactor for Large Bone Tissue Engineering Scaffolds. Appl Biochem Biotechnol 2018; 185(2): 555-563.
  • [26] Porter B, Zauel R, Stockman H, Guldberg R, Fyhrie D. 3-D computational modeling of media flow through scaffolds in a perfusion bioreactor. J Biomech 2005; 38: 543–549.
  • [27] Lamalice L, Le Boeuf F, Huot J. Endothelial cell migration during angiogenesis. Circ Res 2007; 100: 782-794.
  • [28] Li S, Huang NF, Hsu S. Mechanotransduction in endothelial cell migration. J Cell Biochem 2005; 96: 1110–1126.
  • [29] Rennier K, Ji JY. Shear stress regulates expression of death-associated protein kinase in suppressing TNFα-induced endothelial apoptosis. J Cell Physiol 2012; 227: 2398-2411.
  • [30] Chou PH, Wang ST, Yen MH, Liu CL, Chang MC, Lee OK. Fluid-induced, shear stress-regulated extracellular matrix and matrix metalloproteinase genes expression on human annulus fibrosus cells. Stem Cell Res Ther 2016; 7: 34.
  • [31] Asahara T, Takahashi T, Masuda H, Kalka C, Chen D, Iwaguro H, Inai Y, Silver M, Isner JM. VEGF contributes to postnatal neovascularization by mobilizing bone marrow-derived endothelial progenitor cells. EMBO J 1999; 18: 3964-3972.
  • [32] Bao S, Wu Q, Sathornsumetee S, Hao Y, Li Z, Hjelmeland AB, Shi Q, McLendon RE, Bigner DD, Rich JN. Stem cell-like glioma cells promote tumor angiogenesis through vascular endothelial growth factor. Cancer Res 2006; 66: 7843-7848.
  • [33] Beckermann BM, Kallifatidis G, Groth A, Frommhold D, Apel A, Mattern J, Salnikov AV, Moldenhauer G, Wagner W, Diehlmann A, et al. VEGF expression by mesenchymal stem cells contributes to angiogenesis in pancreatic carcinoma. Br J Cancer 2008; 99: 622-631.
  • [34] Cheng BB, Yan ZQ, Yao QP, Shen BR, Wang JY, Gao LZ, Li YQ, Yuan HT, Qi YX, Jiang ZL. Association of SIRT1 expression with shear stress induced endothelial progenitor cell differentiation. J Cell Biochem 2012; 113(12): 3663-3671.
  • [35] Yang Z, Xia WH, Zhang YY, Xu SY, Liu X, Zhang XY, Yu BB, Qiu YX, Tao J. Shear stress-induced activation of Tie2-dependent signaling pathway enhances reendothelialization capacity of early endothelial progenitor cells. J Mol Cell Cardiol 2012; 52(5): 1155-1163.
  • [36] Ohtani-Kaneko R, Sato K, Tsutiya A, Nakagawa Y, Hashizume K, Tazawa H. Characterisation of human induced pluripotent stem cell-derived endothelial cells under shear stress using an easy-to-use microfluidic cell culture system. Biomed Microdevices 2017; 19(4): 91.
  • [37] Chen WT, Hsu WT, Yen MH, Changou CA, Han CL, Chen YJ, Cheng JY, Chang TH, Lee OK, Ho JH. Alteration of mesenchymal stem cells polarity by laminar shear stimulation promoting β-catenin nuclear localization. Biomaterials 2019; 190-191: 1-10.
  • [38] Laschke MW, Harder Y, Amon M, Martin I, Farhadi J, Ring A, Torio-Padron N, Schramm R, Rücker M, Junker D, et al. Angiogenesis in tissue engineering: breathing life into constructed tissue substitutes. Tissue Eng 2006; 12: 2093-2104.
  • [39] Yamamoto K., Takahashi T, Asahara T, Ohura N, Sokabe T, Kamiya A, Ando J. Proliferation, differentiation, and tube formation by endothelial progenitor cells in response to shear stress. J Appl Physiol 2003; 95: 2081-2088.
  • [40] Wang H, Riha GM, Yan S, Li M, Chai H, Yang H, Yao Q, Chen C. Shear stress induces endothelial differentiation from a murine embryonic mesenchymal progenitor cell line. Arterioscler Thromb Vasc Biol 2005; 25: 1817-1823.
  • [41] Park JS, Huang NF, Kurpinski KT, Patel S, Hsu S, Li S. Mechanobiology of mesenchymal stem cells and their use in cardiovascular repair. Front Biosci 2007; 12: 5098-5116.
  • [42] Cui X, Zhang X, Guan X, Li H, Li X, Lu H, Cheng M. Shear stress augments the endothelial cell differentiation marker expression in late EPCs by upregulating integrins. Biochem Biophys Res Commun 2012; 425(2): 419-425.
  • [43] Levy AP, Levy NS, Goldberg MA. Post-transcriptional regulation of vascular endothelial growth factor by hypoxia. J Biol Chem 1996; 271: 2746-2753.
  • [44] Johnson BD, Mather KJ, Wallace JP. Mechanotransduction of shear in the endothelium: basic studies and clinical implications. Vasc Med 2011; 16(5): 365-377.

THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR

Yıl 2019, , 179 - 190, 31.07.2019
https://doi.org/10.18036/estubtdc.598774

Öz



ABSTRACT



 



Packed-bed bioreactors provide
larger surface area to volume ratio compared to the static culture on flasks.
Therefore, these systems offer ideal production environment for large-scale
culture of mesenchymal stem cells (MSCs), but the effect of fluid dynamics on
the cell-behavior of MSCs is not fully elucidated. In this study, packed-bed
perfusion reactor loaded with different size of polymethyl methacrylate
carriers was used to apply different rates of shear stress and drug forces at
constant flow rate. The cell viability, cell-expansion, apoptosis and protein
secretion levels were analyzed for both unmodified and Vascular Endothelial
Growth Factor-positive (VEGF+) MSCs. The superficial stress was
estimated to between 0.21-0.25 N/m2. The results showed that the
shear stress reduced the VEGF secretion, and Caspase-3 was activated at high
drag force, which cause the reduction of the cell numbers in the bioreactor.
The reduction of cytoskeletal actin structures seemed to play the central role
in this adverse effect of the non-planar shear stress.  The expression reduction of VEGF might also
have critical impacts on the tissue engineering applications, in which the
formation of vascular construct is essential.



 

Kaynakça

  • [1] Zhang ZY, Teoh SH, Teo EY, Khoon Chong MS, Shin CW, Tien FT, Choolani MA, Chan JK. A comparison of bioreactors for culture of fetal mesenchymal stem cells for bone tissue engineering. Biomaterials 2010; 31: 8684-8695.
  • [2] Bancroft GN, Sikavitsas VI, Mikos AG. Design of a flow perfusion bioreactor system for bone tissue-engineering applications. Tissue Eng 2003; 9: 549-554.
  • [3] Kavlock KD, Goldstein AS. Effect of pulse frequency on the osteogenic differentiation of mesenchymal stem cells in a pulsatile perfusion bioreactor. J Biomech Eng 2011; 133: 091005.
  • [4] Sikavitsas VI, Bancroft GN, Holtorf HL, Jansen JA, Mikos AG. Mineralized matrix deposition by marrow stromal osteoblasts in 3D perfusion culture increases with increasing fluid shear forces. Proc Natl Acad Sci USA 2003; 100: 14683–14688.
  • [5] Osiecki MJ, McElwain SDL, Lott WB. Modelling mesenchymal stromal cell growth in a packed bed bioreactor with a gas permeable wall. PLoS One 2018; 13: e0202079.
  • [6] Zhao F, Chella R, Ma T. Effects of shear stress on 3-D human mesenchymal stem cell construct development in a perfusion bioreactor system: Experiments and hydrodynamic modeling. Biotechnol Bioeng 2007; 96: 584-595.
  • [7] Le Blanc K, Davies LC. MSCs-cells with many sides. Cytotherapy 2018; 20: 273-278.
  • [8] Guo T, Yu L, Lim CG, Goodley AS, Xiao X, Placone JK, Ferlin KM, Nguyen BN, Hsieh AH, Fisher JP. Effect of Dynamic Culture and Periodic Compression on Human Mesenchymal Stem Cell Proliferation and Chondrogenesis. Ann Biomed Eng 2016; 44: 2103-2113.
  • [9] Caplan AI. Adult mesenchymal stem cells for tissue engineering versus regenerative medicine. J Cell Physiol 2007; 213: 341–347.
  • [10] Knippenberg M, Helder MN, Doulabi BZ, Semeins CM, Wuisman PI, Klein-Nulend J. Adipose tissue-derived mesenchymal stem cells acquire bone cell-like responsiveness to fluid shear stress on osteogenic stimulation. Tissue Eng 2005; 11: 1780-1788.
  • [11] Huang Y, Jia X, Bai K, Gong X, Fan Y. Effect of fluid shear stress on cardiomyogenic differentiation of rat bone marrow mesenchymal stem cells. Arch Med Res 2010; 41: 497-505.
  • [12] Ghadirian E., Arastoopour H. CFD simulation of a fluidized bed using the EMMS approach for the gas-solid drag force. Powder Technology 2016; 288: 35-44.
  • [13] Wiklund K, Zhang H, Stangner T, Singh B, Bullitt E, Andersson M. A drag force interpolation model for capsule-shaped cells in fluid flows near a surface. Microbiology. 2018;164(4):483-494.
  • [14] Turac G, Duruksu G, Karaoz E. The Effect of Recombinant Tyrosine Hydroxylase Expression on the Neurogenic Differentiation Potency of Mesenchymal Stem Cells. Neurospine 2018; 15: 42-53.
  • [15] Adas G, Percem A, Adas M, Kemik O, Arikan S, Ustek D, Cakiris A, Abaci N, Kemik AS, Kamali G, et al. VEGF-A and FGF gene therapy accelerate healing of ischemic colonic anastomoses (experimental study). Int J Surg 2011; 9: 467-471.
  • [16] Geankoplis CJ. Transport Processes and Unit Operations. 3rd ed. Englewood Cliffs, NJ, USA: Prentice Hall Press, 1993.
  • [17] Bueno EM, Ruberti JW. Optimizing Collagen Transport Through Track-Etched Nanopores.J Memb Sci 2008; 321: 250-263.
  • [18] Matsuura T. Bioreactors for 3-dimensional high-density culture of human cells. Hum Cell 2006; 19: 11-16.
  • [19] Mitra D, Whitehead J, Yasui OW, Leach JK. Bioreactor culture duration of engineered constructs influences bone formation by mesenchymal stem cells. Biomaterials 2017; 146: 29-39.
  • [20] Li YJ, Batra NN, You LD, Meier SC, Coe IA, Yellowley CE, Jacobs CR. Oscillatory fluid flow affects human marrow stromal cell proliferation and differentiation. J Orthop Res 2004; 22: 1283–1289.
  • [21] Hosseinkhani H, Inatsugu Y, Hiraoka Y, Inoue S, Tabata Y. Perfusion culture enhances osteogenic differentiation of rat mesenchymal stem cells in collagen sponge reinforced with poly(glycolic acid) fiber. Tissue Eng 2005; 11: 1476–1488.
  • [22] Yourek G, McCormick SM, Mao JJ, Reilly GC. Shear stress induces osteogenic differentiation of human mesenchymal stem cells. Regen Med 2010; 5: 713-724.
  • [23] Kim SH, Ahn K, Park JY. Responses of human adipose-derived stem cells to interstitial level of extremely low shear flows regarding differentiation, morphology, and proliferation. Lab Chip 2017; 17(12): 2115-2124.
  • [24] Dong JD, Gu YQ, Li CM, Wang CR, Feng ZG, Qiu RX, Chen B, Li JX, Zhang SW, Wang ZG, et al. Response of mesenchymal stem cells to shear stress in tissue-engineered vascular grafts. Acta Pharmacol Sin 2009; 30: 530-536.
  • [25] Bhaskar B, Owen R, Bahmaee H, Rao PS, Reilly GC. Design and Assessment of a Dynamic Perfusion Bioreactor for Large Bone Tissue Engineering Scaffolds. Appl Biochem Biotechnol 2018; 185(2): 555-563.
  • [26] Porter B, Zauel R, Stockman H, Guldberg R, Fyhrie D. 3-D computational modeling of media flow through scaffolds in a perfusion bioreactor. J Biomech 2005; 38: 543–549.
  • [27] Lamalice L, Le Boeuf F, Huot J. Endothelial cell migration during angiogenesis. Circ Res 2007; 100: 782-794.
  • [28] Li S, Huang NF, Hsu S. Mechanotransduction in endothelial cell migration. J Cell Biochem 2005; 96: 1110–1126.
  • [29] Rennier K, Ji JY. Shear stress regulates expression of death-associated protein kinase in suppressing TNFα-induced endothelial apoptosis. J Cell Physiol 2012; 227: 2398-2411.
  • [30] Chou PH, Wang ST, Yen MH, Liu CL, Chang MC, Lee OK. Fluid-induced, shear stress-regulated extracellular matrix and matrix metalloproteinase genes expression on human annulus fibrosus cells. Stem Cell Res Ther 2016; 7: 34.
  • [31] Asahara T, Takahashi T, Masuda H, Kalka C, Chen D, Iwaguro H, Inai Y, Silver M, Isner JM. VEGF contributes to postnatal neovascularization by mobilizing bone marrow-derived endothelial progenitor cells. EMBO J 1999; 18: 3964-3972.
  • [32] Bao S, Wu Q, Sathornsumetee S, Hao Y, Li Z, Hjelmeland AB, Shi Q, McLendon RE, Bigner DD, Rich JN. Stem cell-like glioma cells promote tumor angiogenesis through vascular endothelial growth factor. Cancer Res 2006; 66: 7843-7848.
  • [33] Beckermann BM, Kallifatidis G, Groth A, Frommhold D, Apel A, Mattern J, Salnikov AV, Moldenhauer G, Wagner W, Diehlmann A, et al. VEGF expression by mesenchymal stem cells contributes to angiogenesis in pancreatic carcinoma. Br J Cancer 2008; 99: 622-631.
  • [34] Cheng BB, Yan ZQ, Yao QP, Shen BR, Wang JY, Gao LZ, Li YQ, Yuan HT, Qi YX, Jiang ZL. Association of SIRT1 expression with shear stress induced endothelial progenitor cell differentiation. J Cell Biochem 2012; 113(12): 3663-3671.
  • [35] Yang Z, Xia WH, Zhang YY, Xu SY, Liu X, Zhang XY, Yu BB, Qiu YX, Tao J. Shear stress-induced activation of Tie2-dependent signaling pathway enhances reendothelialization capacity of early endothelial progenitor cells. J Mol Cell Cardiol 2012; 52(5): 1155-1163.
  • [36] Ohtani-Kaneko R, Sato K, Tsutiya A, Nakagawa Y, Hashizume K, Tazawa H. Characterisation of human induced pluripotent stem cell-derived endothelial cells under shear stress using an easy-to-use microfluidic cell culture system. Biomed Microdevices 2017; 19(4): 91.
  • [37] Chen WT, Hsu WT, Yen MH, Changou CA, Han CL, Chen YJ, Cheng JY, Chang TH, Lee OK, Ho JH. Alteration of mesenchymal stem cells polarity by laminar shear stimulation promoting β-catenin nuclear localization. Biomaterials 2019; 190-191: 1-10.
  • [38] Laschke MW, Harder Y, Amon M, Martin I, Farhadi J, Ring A, Torio-Padron N, Schramm R, Rücker M, Junker D, et al. Angiogenesis in tissue engineering: breathing life into constructed tissue substitutes. Tissue Eng 2006; 12: 2093-2104.
  • [39] Yamamoto K., Takahashi T, Asahara T, Ohura N, Sokabe T, Kamiya A, Ando J. Proliferation, differentiation, and tube formation by endothelial progenitor cells in response to shear stress. J Appl Physiol 2003; 95: 2081-2088.
  • [40] Wang H, Riha GM, Yan S, Li M, Chai H, Yang H, Yao Q, Chen C. Shear stress induces endothelial differentiation from a murine embryonic mesenchymal progenitor cell line. Arterioscler Thromb Vasc Biol 2005; 25: 1817-1823.
  • [41] Park JS, Huang NF, Kurpinski KT, Patel S, Hsu S, Li S. Mechanobiology of mesenchymal stem cells and their use in cardiovascular repair. Front Biosci 2007; 12: 5098-5116.
  • [42] Cui X, Zhang X, Guan X, Li H, Li X, Lu H, Cheng M. Shear stress augments the endothelial cell differentiation marker expression in late EPCs by upregulating integrins. Biochem Biophys Res Commun 2012; 425(2): 419-425.
  • [43] Levy AP, Levy NS, Goldberg MA. Post-transcriptional regulation of vascular endothelial growth factor by hypoxia. J Biol Chem 1996; 271: 2746-2753.
  • [44] Johnson BD, Mather KJ, Wallace JP. Mechanotransduction of shear in the endothelium: basic studies and clinical implications. Vasc Med 2011; 16(5): 365-377.
Toplam 44 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Yapısal Biyoloji
Bölüm Makaleler
Yazarlar

Gökhan Duruksu 0000-0002-3830-2384

Yayımlanma Tarihi 31 Temmuz 2019
Yayımlandığı Sayı Yıl 2019

Kaynak Göster

APA Duruksu, G. (2019). THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR. Eskişehir Teknik Üniversitesi Bilim Ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji, 8(2), 179-190. https://doi.org/10.18036/estubtdc.598774
AMA Duruksu G. THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR. Eskişehir Teknik Üniversitesi Bilim ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji. Temmuz 2019;8(2):179-190. doi:10.18036/estubtdc.598774
Chicago Duruksu, Gökhan. “THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR”. Eskişehir Teknik Üniversitesi Bilim Ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji 8, sy. 2 (Temmuz 2019): 179-90. https://doi.org/10.18036/estubtdc.598774.
EndNote Duruksu G (01 Temmuz 2019) THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR. Eskişehir Teknik Üniversitesi Bilim ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji 8 2 179–190.
IEEE G. Duruksu, “THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR”, Eskişehir Teknik Üniversitesi Bilim ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji, c. 8, sy. 2, ss. 179–190, 2019, doi: 10.18036/estubtdc.598774.
ISNAD Duruksu, Gökhan. “THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR”. Eskişehir Teknik Üniversitesi Bilim ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji 8/2 (Temmuz 2019), 179-190. https://doi.org/10.18036/estubtdc.598774.
JAMA Duruksu G. THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR. Eskişehir Teknik Üniversitesi Bilim ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji. 2019;8:179–190.
MLA Duruksu, Gökhan. “THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR”. Eskişehir Teknik Üniversitesi Bilim Ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji, c. 8, sy. 2, 2019, ss. 179-90, doi:10.18036/estubtdc.598774.
Vancouver Duruksu G. THE EFFECT OF DRAG FORCE AND FLOW RATE ON MESENCHYMAL STEM CELLS IN PACKED-BED PERFUSION BIOREACTOR. Eskişehir Teknik Üniversitesi Bilim ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji. 2019;8(2):179-90.