مقایسه پتانسیل تکثیری سلولهای بنیادی مشتق از بافت چربی ناحیه دنبه، اینفرا پاتلا و زیر پوستی در مدل حیوانی گوسفند

نویسندگان

1 گروه علوم تشریح، دانشکده علوم پزشکی، دانشگاه علوم پزشکی، مراغه، ایران

2 مرکز تحقیقات، اداره دامپزشکی، مراغه، ایران

3 گروه علوم تشریح، دانشکده علوم پزشکی، دانشگاه علوم پزشکی، تبریز، ایران

چکیده

مقدمه و هدف: سلولهای بنیادی از منابع مختلفی به دست می آیند و به دلیل نقش آنها در درمان و بهبود بافت های آسیب دیده شناسایی منابع سلولهای بنیادی و بررسی آنها از نظر پژوهشگران بسیار با اهمیت می باشد، لذا هدف از این مطالعه شناسایی منابع بافتی ایده آل و به دست آوردن سلولهای بنیادی با پتانسیل تکثیری و تمایزی مناسب می باشد.
مواد و روش ها: نمونه بافت در کشتارگاه از ناحیه دنبه، زیر جلدی و اینفراپاتلای گوسفند نر با سن 2 ساله برداشته شد و نمونه ها در آزمایشگاه تکه تکه شدند و تحت تأثیر آنزیم کلاژناز قرار گرفتند؛ و سلولها با لام نئوبار زیر میکروسکوپ اینورت شمارش شدند و به تعداد 105 سلول به هر فلاسک کشت داده شدند و برای تعیین مدت دو برابر شدن سلولها از فرمول پترسون Td=Tlg2/lg(Nt/N0)  استفاده شد. داده ها با استفاده از آنالیز واریانس تحلیل شدند (05/0P>).
نتایج: یافته ها نشان دادند که سلولهای بنیادی به دست آمده از هر سه ناحیه در پاساژهای اولیه توان تکثیری و مورفولوژی مشابه به هم داشتند ولی در پاساژهای بالاتر، سلولهای به دست آمده از بافت چربی ناحیه اینفراپاتلا در مقایسه با سلولهای زیرپوستی و دنبه در مدت زمان کمتری به ظرفیت تکثیری 80%(confluenc) رسیدند.
نتیجه‌گیری: نتیجه‌گیری می شود که سلولهای بنیادی ناحیه ایفرا پاتلا در مقایسه با سلولهای بافت چربی سایر نواحی، برای مدت طولانی پتانسیل تکثیری خودشان را حفظ می‌کنند لذا بافت چربی ناحیه اینفرا پاتلا یک منبع ایده ال برای تهیه سلولهای بنیادی می تواند باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Comparative study of the proliferation potency of adipose tissue derived stem cells from infrapatellar area, subcutaneous adipose tissue and tail fat pad in a sheep animal model

نویسندگان [English]

  • Parviz Vahedi 1
  • Masood Madadi 2
  • Ahad Ferdoosi khosroshahi 3
چکیده [English]

Background and Objective: The stem cells are obtained from different sources and due to the role of them in the treatment and recovery of damaged tissues, characterizing and identifying of the sources of stem cells and investigating them is particularly interesting for researchers. Thus, the purpose of this study was to identify ideal tissue sources and capture stem cells with appropriate differentiation and proliferation potential.
Materials and Methods: The adipose tissue sample were harvested from the tail adipose tissue, subcutaneous and infrapatellar in the slaughterhouse from male sheep about 2 years old and were rinsed to small pieces in the laboratory and were treated by the collagenase enzyme and the cells were counted by Neubauer chamber with inverted microscope and then about 105 cells were cultured in T25 flask and was calculated the doubling time by deleted the Peterson formula Td=Tlg2/lg(Nt/N0). Data were analyzed using analysis of variance (P< 0/02).
Results: Our findings showed that the stem cells of the three areas in the early passages are similar to each other in regarding proliferation and morphological potentials, but in higher passages as compared to the subcutaneous and tail fat, the cells of the infrapatellar region received confluence 80% in shorter time than the others.
Conclusion: The results of this study showed that infrapatellar adipose tissue stem cells maintain their proliferation potential and differentiation for long time as compared to adipose tissue of other areas. So infrapatellar fat pad can be an ideal source for obtaining stem cells.
 

کلیدواژه‌ها [English]

  • Stem cells
  • Subcutaneous adipose tissue stem cells
  • Cell proliferation
  • Infrapatellar fat pad stem cells
1. Pinar Yilgor H, Seren Ha, Emre E, Gazi H, Mahmut N. Infrapatellar Fat Pad-Derived Stem Cell-Based Regenerative Strategies in Orthopedic Surgery. Knee Surgery & Related Research 2018;30(3):179-186. 2. Woo DH, Hwang HS, Shim JH. Comparison of adult stem cells derived from multiple stem cell niches. Biotechnology Letters 2016;38:751–759. 3. Weiping L, Linfeng H, Ying Li, Bin F, Gang Li, Leilei Ch, Liangliang Xu. Mesenchymal Stem Cells and Cancer:Clinical Challenges and Opportunities. BioMed Research International 2019; 7: 2820-853. 4. Koobatian MT, Liang MS, Swartz DD, Andreadis ST. Differential effects of culture senescence and mechanical stimulation on the proliferation and leiomyogenic differentiation of MSC from different sources: implications for engineering vascular grafts. Tissue Engineering, Parts A 2015; 21:1364–1375. 5. Hu YL, Fu YH, Tabata Y, Gao JQ. Mesenchymalstem cells: A promising targeted-delivery vehicle in cancer gene therapy. Journal of Controlled Release 2010; 147:154-16. 6. Zhao L, Johnson T, Liu D. Therapeutic angiogenesis of adipose-derived stem cells for ischemic diseases. Stem Cell Research Therapy 2017; 8: 125. 7. Naderi N, Combellack EJ, Gri_nM, Sedaghati T, Javed M, et al. The regenerative role of adipose- derived stem cells (ADSC) in plastic and reconstructive surgery. International Wound Journal 2017; 14: 112–124. 8. Hirano A, Sano M, Urushihata N, Tanemura H, Oki K, Suzaki E. Assessment of safety and feasibility of human allogeneic adipose-derived mesenchymal stem cells in a pediatric patient. Pediatric Research 2018, 84: 575–577. 9. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI et al. Human adipose tissue is a source of multipotent stem cells. Molecular Biology of the Cell 2002; 13: 4279–4295. 10. Lopa S, Colombini A, Stanco D et al. Donor-matched mesenchymal stem cells from knee infrapatellar and subcutaneous adipose tissue of osteoarthritic donors display differential chondrogenic and osteogenic commitment. European Cells & Materials 2014; 27: 298-311. 11. Stefan A, Sabine W. Adipose tissue derived mesenchymal stem cells for musculoskeletal repair in veterinary medicine. American Journal of Stem Cells 2015;4(1):1-12. 12. Chu DT, Tao Y, Son LH, Le DH et al. Cell source, differentiation, functional stimulation, and potential application of human thermogenic adipocytes in vitro. Journal of Physiology and Biochemistry 2016, 73, 315–321. 13. Ding DC, Wu KC, Chou HL, et al. Human infrapatellar fat pad-derived stromal cells have more potent differentiation capacity than other mesenchymal cells and can be enhanced by hyaluronan. Cell Transplant 2015; 24: 1221- 32. 14. Hindle P, Khan N, Biant L, Pe´ ault B. The infrapatellar fat pad as a source of perivascular stem cells with increased chondrogenic potential for regenerative medicine. Stem Cells Transplation Medicine 2017; 6: 77- 87. 15. Siciliano C, Bordin A, Ibrahim M, Chimenti I, Cassiano F, Gatto I, et al. The adipose tissue of origin influences the biological potential of human adipose stromal cells isolated from mediastinal and subcutaneousfat depots. Stem Cell Research 2016; 17: 342-51. 16. Dominik DU, Anna L, Robert Re, David A, Zeshaan N. Suction assisted liposuction does not impair the regenerative potential of adipose derived stem cells. Journal of Translational Medicine 2016;12: 118- 126. 17. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, et al. Multilineage cells from human adipose tissue: Implications for cell-based therapies. In: Advances in Tissue Engineering: Stem Cells and Developments 2010; 23: 119-33. 18. Ribeiro A, Laranjeira P, Mendes S, Martinho A, Pais M, et al. Mesenchymal stem cells from umbilical cord matrix, adipose tissue and bone marrow exhibit different capability to suppress peripheral blood B, natural killer and T cells. Stem Cell Ressearch Therapy 2013. 19. Pizzute T, Lynch K, Pei M. Impact of tissue-specific stem cells on lineage specific differentiation: a focus on musculoskeletal system. Stem Cell Reviews and Reports 2015;11:119- 32. 20. Wei X, Peng G, Zheng S. Differentiation of umbilical cord mesenchymal stem cells into steroidogenic cells in comparison to bone marrow mesenchymal stem cells. Cell Proliferation 2012; 45: 101-10. 21. Sun Yu, Song Ch and Ming P. Comparative advantages of infrapatellar fat pad: an emerging stem cell source for regenerative medicine. Rheumatology 2018; 57:2072 -2086. 22. Somoza RA, Welter JF, Correa D, Caplan AI. Chondrogenic differentiation of mesenchymal stem cells: challenges and unfulfilled expectations. Tissue Engineering Part B Review. 2014; 20:596- 608. 23. Ronaldo J, Henrique V, Almeid A, Daniel J, Fergal J, et al. Infrapatellar Fat Pad Stem Cells: From Developmental Biology to Cell Therapy. Stem Cells International 2017, 30(3): 179–186. 24. Dinh Ch, Thuy N, Nguyen Le, Bao T, Dang K, Adipose Tissue Stem Cells for Therapy: An Update on the Progress of Isolation, Culture, Storage, and Clinical Application. Journal Clinical Medicine 2019; 8: 917-936. 25. Si Z, Wang X, Sun C, Kang Y, Xu J, Wang X, et al. Adipose-derived stem cells: Sources, potency, and implications for regenerative therapies. Biomed. Pharmacother 2019; 114: 108 -130.