HEMATOPOETİK KÖK HÜCRE İZOLASYONUNDA GÜNCEL YÖNTEMLER

Hematopoetik kök hücreler (HKH) başta kemik iliği (Kİ) olmak üzere çeşitli hematopoetik organlarda, kordon kanında (KK), aferez materyalinde ve periferik kanda çok az sayıda bulunmaktadır (1-100 hücre/ml). Bu hücrelerin saflaştırılarak diyabet, kanser, kalp, nörolojik hastalıklar, kas ve eklem sorunları olan hastalara nakledilmesi fonksiyonu bozulan doku ve organların rejenerasyonunu sağlayacaktır. Kanser hastalarında otolog kök hücrelerin transplantasyonunda en önemli sorun malign hücre kontaminasyonudur. Allojenik transplantasyonda ise hücreler farklı kişilerden alındığı için, alıcıda meydana gelen immün reaksiyonlar ve doku reddi bu alanın en önemli sorunudur. Dolayısıyla transplant materyalinin tümör hücrelerinden ve immün hücrelerden temizlenmesi nakledildiği dokuda uzun süreli ve sağlıklı rejenerasyonu sağlayacaktır. Klinik kullanım için bu hücrelerin saflaştırılmasında uzun yıllar çeşitli teknikler geliştirilmesine rağmen halen uygun ve etkili bir yöntem bulunmamaktadır. Son yıllarda geliştirilen mikroakışkan sistemler (mikroçipler) kolay, ucuz ve yüksek verimli olması nedeni ile araştırılmaktadır. Çeşitli mikrokapiller, mikrosütunlar, mikroporlarla dizayn edilen bu sistemlerde hücrelerin boyut, mekanik esneklik, deformabilite, hücre adezyonu ve elektrik yükleri gibi özelliklerine dayalı saflaştırma yapılmaktadır. Başlangıçta çok az sayıda bulunan dolaşımdaki tümör hücrelerini (DTH) yakalamak amacıyla geliştirilen bu sistemler şimdi çeşitli hücrelerin, mikroveziküllerin ve eksozomların saflaştırılması ve tanısı için kullanılmaktadır. Bu derleme, geleneksel ve gelişmekte olan hematopoetik kök hücre izolasyon yöntemleri ile bu yöntemlerin avantajları ve dezavantajlarını açıklamayı amaçlamaktadır.

Anahtar Kelimeler:

Kök hücre izolasyonu, mikroakışkan sistemler, .

CURRENT METHODS IN ISOLATION OF HEMATOPOIETIC STEM CELL

Hematopoietic stem cells are the rare cells in various hematopoietic organs which are found in small numbers (1-100/ml). When these cells are transferred to patients suffering from diabetes, cancer, heart diseases, muscle and joint problems, they will help the patients’ bodies with the regeneration of impaired tissues. While the most important problem in the transplantation of autologous stem cells is malignant cell contamination in cancer patients, in allogeneic transplantation, it is immune reactions and tissue rejections. Therefore, clearing of the transplant material from tumour cells and immune cells may support the long-term healthy regeneration of the tissue to which they are transplanted. Although various techniques have been developed for the purification of these cells in terms of clinical use over many years, there is still no sufficiently effective method. In recent years, researchers have shown an increased interest in microfluidic systems because they are easy to use, cheap and highly efficient. In these types of systems designed with various microcapillaries, micropillars and micropores; purification is carried out according to the properties of cells such as size, deformability, cell adhesion and electrical charges. This review aims to explain traditional and emerging hematopoietic stem cell isolation methods and their advantages and disadvantages.

Keywords:

Stem cell isolation, microfluidic system,

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1. Gou Y, Jia Y, Wang P, Sun C. Progress of ınertial microfluidics in principle and application. Sensors (Basel) 2018;18(6):1762. [CrossRef]
2. Tomlinson MJ, Tomlinson S, Yang, XB, Kirkham K. Cell separation: Terminology and practical considerations. Journal of Tissue Engineering 2012; 3(1):1-14.
3. Metin S, Dere H. Hematopoetik kök hücre nakli ve güncel beslenme yaklaşımları. Turkiye Klinikleri J Pediatr 2017;26(1):22-31. [CrossRef]
4. Zhu B, Murthy SK. Stem cell separation technologies. Curr Opin Chem Eng. 2014;2(1):3-7. [CrossRef]
5. Rahmanian N, Bozorgmehr M, Torabi M, Akbari A, Zarnani AH. Cell separation: Potentials and pitfalls. Prep Biochem Biotechnol 2017;47(1):38-51. [CrossRef]
6. Kenry, Leed WC, Loh KP, Li CT. When stem cells meet graphene: Opportunities and challenges inregenerative medicine. Biomaterials 2018;155:236-50. [CrossRef]
7. Erdem-Kuruca S, Celik DD, Demirel G, Ozerkan D. Characterization and isolation of very small embryonic-like (VSEL) stem cells obtained from human peripheral blood. Stem Cell Rev Rep 2019;15(5):730-42. [CrossRef]
8. Chhabra P, Brayman KL. Stem cell therapy to cure type 1 diabetes: from hype to hope. Stem Cells Transl Med 2013;2(5):328-36. [CrossRef]
9. Mazini L, Rochette L, Amine M, Malka, G. Regenerative capacity of adipose derived stem cells (adscs), comparison with mesenchymal stem cells (mscs). Int J Mol Sci 2019;20(10):2523. [CrossRef]
10. Zhang CL, Huang T, Wu BL, He WX, Liu D. Stem cells in cancer therapy: opportunities and challenges. Oncotarget 2017;8(43):75756-66. [CrossRef]
11. Almendros I, Carreras A, Montserrat JM., Gozal D, Navajas D, Farre R. Potential role of adult stem cells in obstructive sleep apnea. Front Neurol 2012;3:112. [CrossRef]
12. Yo M, Sakaue-Sawano A, Noda S, Miyawaki A, Miyoshi H. Fucci-guided purification of hematopoietic stem cells with high repopulating activity. Biochem Biophys Res Commun 2015;30;457(1):7-11. [CrossRef]
13. Ng AP, Alexander WS. Haematopoietic stem cells: past, present and future. Cell Death Discovery 2017;3:17002. [CrossRef]
14. Sargın D. Kök hücre ve kök hücre tedavisi. XXX. Ulusal Hematoloji Kongresi Mezuniyet Sonrası Eğitim Kursu; 10-14 Ekim 2003; İstanbul, 2003. p. 49-61.
15. Diogo MM, Silva CL, Cabral JMS. Chapter 7: Separation technologies for stem cell bioprocessing. In: Mohamed Al-Rubeai, Mariam Naciri, editors. Stem Cells and Cell Therapy. Dordrecht: Springer; 2014. p. 157-181. [CrossRef]
16. Hoeve MA, De Sousa, PA, Willoughby NA. Chapter: 5. Challenges of scale-up of cell separation and purification techniques: 5.3.3.1. Centrifugal Counterflow Elutriation (CCE). In: Connon Che J, editor. Bioprocessing for Cell- Based Therapies. UK: John Wiley & Sons; 2017. p. 138-140.
17. Bachère E, Chagot D, Henrı Grızel H. Cell Separation by centrifugal elutriation. American Fisheries Society Special Publication 1988;18:281-5.
18. Higuchi A, Lizuka A, Gomei Y, Miyazaki T, Sakurai M, Matsuoka Y, et al. Separation of CD34+ cells from human peripheral blood through polyurethane foaming membranes. J Biomed Mater Res A 2006;78(3):491-9. [CrossRef]
19. Muller-Steinhardt M, Hennig H, Kirchner H, Schlenke P. Prestorage WBC filtration of RBC units with soft-shell filters: Filtration performance and impact on RBCs during storage for 42 days. Transfusion 2002;42:153-8. [CrossRef]
20. Fatanat T, Li K, Veres T, Tabrizian M. Separation of rare oligodendrocyte progenitor cells from brain using a highthroughput multilayer thermoplastic-based micro ﬂuidic device. Biomaterials 2013;34:5588-93. [CrossRef]
21. Yousuff CM, Wei-Ho ET, Ismail Hussain K, Hamid NHB. Microﬂuidic platform for cell isolation and manipulation based on cell properties. Micromachines 2017; 8 (15): 1-26. [CrossRef]
22. Actor JK, Elsevier’s Integrated Review Immunology and Microbiology. Second Edition. Elsevier Inc.; 2012. p. 71-9. [CrossRef]
23. Prince H, Arens L, Kleinman S. CD4 and CD8 Subsets Defined by Dual-color Cytofluorometry which Distinguish Symptomatic from Asymptomatic Blood Donors Seropositive for Human Immunodeficiency Virus. Diagn. Clin. Immunol 1986;5(4):188-93.
24. Johnson KW, Dooner M, Quesenberry PJ. Fluorescence activated cell sorting: A window on the stem cell. Curr Pharm Biotechnol 2007;8(3):133-9. [CrossRef]
25. Will B, Steidl U. Multi-parameter fluorescence-activated cell sorting and analysis of stem and progenitor cells in myeloid malignancies. Best Pract Res Clin Haematol 2010;23(3):391- 401. [CrossRef]
26. Curtis MG, Walker B, Denny TN. Flow cytometric methods for prenatal and neonatal diagnosis. J Immuno Methods 2011;363(2):198-209. [CrossRef]
27. Garner DL, Evans KM, Seidel GE. Sex-sorting sperm using flow cytometry/cell sorting. Methods Mol Biol. 2013;927:279-95. [CrossRef]
28. Mahmoud TMS. Selection of non-apoptotic, DNA intact spermatozoa. Groningen: University of Groningen. 2009.
29. Rodrigues GMC, Fernandes TG, Rodrigues CAV, Diogo MM, Cabral JMS. Chapter 9- enrichment and separation technologies for stem cell-based therapies; 9.2.1 Adult stem cells.In: Cabral JMS, Cláudia Lobato da Silva, Chase LG, Diogo MM, editors. Stem Cell Manufacturing. Elsevier; 2016. p. 199-213. [CrossRef]
30. Fernandes TG, Diogo MM, Cabral JMS. Stem cell separation. Stem Cell Bioprocessing For Cellular Therapy, Diagnostics and Drug Development. Woodhead Publishing Series in Biomedicine; 2013. p. 115-41. [CrossRef]
31. Firer MA. Efficient elution of functional proteins in affinity chromatography. J Biochem Biophys Methods 2001;49:433– 42. [CrossRef]
32. Hage DS, Cazes, J, editors. Handbook of Affinity Chromatography. 2nd Edition. Boca Raton: CRC Pres; 2005. p. 856. [CrossRef]
33. Hage DS, Clarke W. Affinity chromatography: In: Cazes J, editör. Encyclopedia of Chromatography. New York: Marcel Dekker; 2004. p. 40-3.
34. Sousa AF, Andrade PZ, Pirzgalska RM, Galhoz TM, Azevedo AM, Silva CL, et al. A novel method for human hematopoietic stem/progenitor cell isolation from umbilical cord blood based on immunoaffinity aqueous two-phase partitioning. Biotechnol Lett 2011;33(12):2373-7. [CrossRef]
35. Yılmaz M. Design and fabrication of low cost passive microfluidic systems for particle separation. Istanbul: Istanbul Technical University, 2012.
36. Kim GY, Han JI, Park JK. Inertial microfluidics-based cell sorting. BioChip Journal 2018;12(4):257-67. [CrossRef]
37. Roda B, Lanzoni G, Alviano F, Zattoni A, Costa R, Di Carlo A, et al. A novel stem cell tag-less sorting method. Stem Cell Rev 2009;5(4):420-7. [CrossRef]
38. Roda B, Reschiglian P, Alviano F, Lanzoni G, Bagnara GP, Ricci F, et al. Gravitational field-flow fractionation of human hemopoietic stem cells. J Chromatogr A 2009;1216(52):9081- 7. [CrossRef]
39. Talary MS, Mills KI, Hoy T, Burnett AK, Pethig R. Dielectrophoretic separation and enrichment of CD34+ cell subpopulation from bone marrow and peripheral blood stem cells. Medical and Biological Engineering and Computing 1995; 33 (2), p. 235-7. [CrossRef]
40. Voldman, J. Electrical forces for microscale cell manipulation. Annu Rev Biomed EnG 2006;8:425-54. [CrossRef]
41. Gwak H, Kim J, Kashefi-Kheyrabadi L, Kwak B, Kyung-A Hyun, Hyo-Il Jung. Progress in circulating tumor cell research using microfluidic devices. micromachines 2018;9(7):353. [CrossRef]
42. Lecault V, Vaninsberghe M, Sekulovic S, Knapp DJ, Wohrer S, Bowden W, et al. High-throughput analysis of single hematopoietic stem cell proliferation in microfluidic cell culture arrays. Nat Methods 2011;8(7):581-6. [CrossRef]
43. Perutelli P, Catellani S, Scarso L, Cornaglia-Ferraris P, Dini G. Processing of human cord blood by three different procedures for red blood cell depletion and mononuclear cell recovery. Vox Sanguinis 1999;76:237-40. [CrossRef]
44. Rocha V, Gluckman E. Eurocord and European Blood and Marrow Transplant Grup. Clinical use of umbilical cord blood hematopoietic stem cells. Biol Blood Marrow Transplant 2006;12(1S):34-41. [CrossRef]
45. Trewhitt KG. Bone marrow aspiration and biopsy: Collection and ınterpretation. Oncol Nurs Forum 2001;28(9):1409-17.
46. Franklin WA, Shpall EJ, Archer P, Johnston CS, Garza- Williams S, Hami L, et al. Immunocytochemical detection of breast cancer cells in marrow and peripheral blood of patients undergoing high dose chemotherapy with autologous stem cell support, Breast Cancer Res Treat 1996;41(1):1-13. [CrossRef]
47. Sharp JG, Kessinger MA, Vaughan WP, Mann SL, Crouse DA, Dicke KA, et al. Detection and clinical significance of minimal tumor cell contamination of peripheral stem cell harverts. Int Cell Cloning 1992;10(1):92-4. [CrossRef]
48. Moss TJ. Sensitive detection of metastatic tumor cells in bone marrow. Prog Clin Biol Res. 1994;389:567-77.
49. Nieto Y, Shpall EJ. CD34+ blood stem cell transplantation. In: Reiffers J, Goldman J, Armitage JO (eds). Blood Stem Cell Transplantation. London: Martin Dunitz Ltd.; 1998. p. 187-201.
50. Ertuğrul-Örüç N, Yenicesu İ. Bölüm-II: Transfüzyon Merkezi. Ulusal Kan ve Kan Bileşenleri Hazırlama, Kullanım ve Kalite Güvencesi Rehberi, Türkiye 2008 Ulusal IPA (Katılım Öncesi Mali Yardım) Programı: 2016. URL: https://www.kanver.org/ Upload/Dosya/ulusal_kan_rehberi. Erişim Tarihi: 07.06.2019.