Effects of fibrin matrix and Ishikawa cells on in vitro 3D uterine tissue cultures on a rat model: A controlled study

Öz

Background/Aim: In recent years, developing an embryo in in vitro conditions has been one of the most challenging and popular objectives in reproductive biology. In vitro models make observing the relationship between the two possible. Various cell culture and matrix models have been created to overcome embryonic disorganization during culturing. The primary aim of this study was to evaluate and compare the effects of fibrin, Ishikawa cell line, and a combination of both on the 3D multilayer uterine tissue cultures on a rat model, including a control group. Methods: This study was designed as a prospective controlled cohort study. The standard uterine culture model [CNT] (N: 3) constituted the control group. In addition, fibrin matrix-supported [FIB] (N: 3), Ishikawa cells-supported [ISH] (N: 3), and a combination of both [FIB+ISH] (N: 3) culture models were designated as the exposures. All models were cultured for 14 days. Afterwards, the optimal model was determined regarding glucose consumption, lactate production, endometrial thickness and gland count (primary outcomes) with semi-quantitative and statistical methods. Finally, the optimal model was implanted with blastocytes, and the survival duration was observed (secondary outcome). Results: There were significant differences between the groups in terms of glucose, lactate, endometrial thickness (millimeter), and the number of endometrial glands (P<0.05). FIB had the least glucose consumption, and the least lactate production was in CNT. The thickest endometrium and most endometrial glands were detected in FIB when all groups were compared, allowing for 14 days of embryo survival. Conclusion: In embryogenesis research, the fibrin-matrix-supported culture model could be a satisfactory 3D uterine tissue culture model.

Anahtar Kelimeler

• Cell line
• Tumor
• Co-culture
• Extracellular matrix
• Fibrin
• In vitro
• Tissue culture techniques

Kaynakça

1. 1. Aguilera-Castrejon A, Oldak B, Shani T, et al. Ex utero mouse embryogenesis from pre-gastrulation to late organogenesis. Nature. 2021 May;593(7857):119-24.
2. 2. Govindasamy N, Duethorn B, Oezgueldez HO, et al. Test-tube embryos - mouse and human development in vitro to blastocyst stage and beyond. Int J Dev Biol. 2019(3-4-5);63:203-15.
3. 3. Deglincerti A, Croft GF, Pietila LN, et al. Self-organisation of the in vitro attached human embryo. Nature. 2016 May 12;533(7602):251-4.
4. 4. Shahbazi MN, Jedrusik A, Vuoristo S, et al. Self-organisation of the human embryo in the absence of maternal tissues. Nat Cell Biol. 2016 Jun;18(6):700-8.
5. 5. Niu Y, Sun N, Li C, et al. Dissecting primate early post-implantation development using long-term in vitro embryo culture. 2019 Nov 15;366(6467):eaaw5754.
6. 6. Nowotschin S, Hadjantonakis AK. Guts and gastrulation: Emergence and convergence of endoderm in the mouse embryo. Curr. Top. Dev. Biol. 2020;136:429-54.
7. 7. Teklenburg G, Weimar CH, Fauser BC, et al. Cell lineage specific distribution of H3K27 trimethylation accumulation in an in vitro model for human implantation. PLoS One. 2012;7(3):e32701.
8. 8. Bentin-Ley U, Horn T, Sjögren A, Sorensen S, Falck Larsen J, Hamberger L. Ultrastructure of human blastocyst-endometrial interactions in vitro. J. Reprod. Fertil. 2000 Nov;120(2):337-50.

9. 9. Wang H, Pilla F, Anderson S, et al. A novel model of human implantation: 3D endometrium-like culture system to study attachment of human trophoblast (Jar) cell spheroids. Mol. Hum. Reprod. 2012 Jan;18(1):33-43.
10. 10. Rozner AE, Durning M, Kropp J, et al. Macrophages modulate the growth and differentiation of rhesus monkey embryonic trophoblasts. Am. J. Reprod. Immunol. 2016 Nov;76(5):364-75.
11. 11. Chang TA, Bondarenko GI, Gerami-Naini B, et al. Trophoblast differentiation, invasion and hormone secretion in a three-dimensional in vitro implantation model with rhesus monkey embryos. Reprod. Biol. Endocrinol. 2018 Mar 16;16(1):24.
12. 12. Geisler K, Künzel J, Grundtner P, Müller A, Beckmann MW, Dittrich R. The perfused swine uterus model: long-term perfusion. Reprod. Biol. Endocrinol. 2012 Dec 15;10:110.
13. 13. Han Y, Biswas D, Yoon JD, Jeon Y, Hyun SH. Effect of porcine uterus as ex vivo model of fertilising ability and gene expression pattern on blastocysts. Theriogenology. 2019 Apr 15;129:146-53.
14. 14. Rossant J. Mouse and human blastocyst-derived stem cells: vive les differences. Development. 2015 Jan 1;142(1):9-12.
15. 15. Rossant J, Tam PPL. New Insights into Early Human Development: Lessons for Stem Cell Derivation and Differentiation. Cell Stem Cell. 2017 Jan 5;20(1):18-28.
16. 16. Shahbazi MN, Zernicka-Goetz M. Deconstructing and reconstructing the mouse and human early embryo. Nat. Cell Biol. 2018 Aug;20(8):878-87.
17. 17. Demir R. Histolojik Boyama Teknikleri. 1st Ed. Ankara: Palme Yayıncılık; 2001.
18. 18. Pathak D, Bansal N, Singh O, et al. Immunohistochemical localisation of estrogen receptor alpha (ERα) in the oviduct of Indian buffalo during follicular and luteal phases of estrous cycle. Trop. Anim. Health Prod. 2019 Jul;51(6):1601-9.
19. 19. Mantikou E, Youssef MA, van Wely M, et al. Embryo culture media and IVF/ICSI success rates: a systematic review. Hum. Reprod. Update. 2013 May-Jun;19(3):210-20.
20. 20. Chan MM, Smith ZD, Grosswendt S, et al. Molecular recording of mammalian embryogenesis. Nature. 2019 Jun;570(7759):77-82.
21. 21. Ibrahim SA, Ackerman WE 4th, Summerfield TL, et al. Inflammatory gene networks in term human decidual cells define a potential signature for cytokine-mediated parturition. Am. J. Obstet. Gynecol. 2016 Feb;214(2):284.e1-284.e47.
22. 22. Moreno D, Neira A, Dubreil L, et al. In vitro bovine embryo production in a synthetic medium: embryo development, cryosurvival, and establishment of pregnancy. Theriogenology. 2015 Oct 15;84(7):1053-60.
23. 23. Block J, Hansen PJ, Loureiro B, Bonilla L. Improving post-transfer survival of bovine embryos produced in vitro: actions of insulin-like growth factor-1, colony stimulating factor-2 and hyaluronan. Theriogenology. 2011 Dec;76(9):1602-9.
24. 24. Pelletier G. Localization of androgen and estrogen receptors in rat and primate tissues. Histopathol. 2000 Oct;15(4):1261-70.
25. 25. Carvalho BR, Barbosa MW, Bonesi H, et al. Embryo stage of development is not decisive for reproductive outcomes in frozen-thawed embryo transfer cycles. JBRA Assist. Reprod. 2017 Feb 1;21(1):23-26.
26. 26. Hannan NJ, Evans J, Salamonsen LA. Alternate roles for immune regulators: establishing endometrial receptivity for implantation. Expert Rev. Clin. Immunol. 2011 Nov;7(6):789-802.
27. 27. Uchida H, Maruyama T, Ohta K, et al. Histone deacetylase inhibitor-induced glycodelin enhances the initial step of implantation. Hum. Reprod. 2007 Oct;22(10):2615-22.
28. 28. Li Y, Moretto-Zita M, Soncin F, et al. BMP4-directed trophoblast differentiation of human embryonic stem cells is mediated through a ΔNp63+ cytotrophoblast stem cell state. Development. 2013 Oct;140(19):3965-76.
29. 29. Jiang X, Li X, Fei X, et al. Endometrial membrane organoids from human embryonic stem cell combined with the 3D Matrigel for endometrium regeneration in asherman syndrome. Bioact. Mater. 2021 Apr 16;6(11):3935-46.
30. 30. Rivron NC, Frias-Aldeguer J, Vrij EJ, et al. Blastocyst-like structures generated solely from stem cells. Nature. 2018 May;557(7703):106-111.
31. 31. van den Brink SC, Baillie-Johnson P, Balayo T, et al. Symmetry breaking, germ layer specification and axial organisation in aggregates of mouse embryonic stem cells. Development. 2014 Nov;141(22):4231-42.
32. 32. Warmflash A, Sorre B, Etoc F, et al. A method to recapitulate early embryonic spatial patterning in human embryonic stem cells. Nat. Methods. 2014 Aug;11(8):847-54.
33. 33. Shao Y, Taniguchi K, Townshend RF, et al. A pluripotent stem cell-based model for post-implantation human amniotic sac development. Nat. Commun. 2017 Aug 8;8(1):208.
34. 34. Zhang D, Lv P, Zhang R, et al. A new model for embryo implantation: coculture of blastocysts and Ishikawa cells. Gynecol. Endocrinol. 2012 Apr;28(4):288-92.
35. 35. Campo H, García-Domínguez X, López-Martínez S, et al. Tissue-specific decellularised endometrial substratum mimicking different physiological conditions influences in vitro embryo development in a rabbit model. Acta Biomater. 2019 Apr 15;89:126-138.
36. 36. Voytik-Harbin SL, Brightman AO, Waisner BZ, et al. Small intestinal submucosa: A tissue-derived extracellular matrix that promotes tissue-specific growth and differentiation of cells in vitro. Tissue engineering. 1998 Jun;4(2):157-74.
37. 37. Saldin LT, Cramer MC, Velankar SS, et al. Extracellular matrix hydrogels from decellularised tissues: Structure and function. Acta Biomater. 2017 Feb;49:1-15.
38. 38. Mühlhauser J, Crescimanno C, Kasper M, et al. Differentiation of human trophoblast populations involves alterations in cytokeratin patterns. J. Histochem. Cytochem. 1995 Jun;43(6):579-89.
39. 39. Kliman HJ, Nestler JE, Sermasi E, et al. Purification, characterisation, and in vitro differentiation of cytotrophoblasts from human term placentae. Endocrinology. 1986 Apr;118(4):1567-82.
40. 40. DaSilva-Arnold S, James JL, Al-Khan A, et al. Differentiation of first trimester cytotrophoblast to extravillous trophoblast involves an epithelialmesenchymal transition. Placenta. 2015 Dec;36(12):1412-8.
41. 41. du Puy L, Lopes SM, Haagsman HP, et al. Analysis of co-expression of OCT4, NANOG and SOX2 in pluripotent cells of the porcine embryo, in vivo and in vitro. Theriogenology. 2011 Feb;75(3):513-26.
42. 42. isaki T, Kawai I, Sugiura K, et al. Regulation of embryonic size in early mouse development in vitro culture system. Zygote. 2014 Aug;22(3):340-7.
43. 43. Enders AC, Meyers S, Vandevoort CA, Douglas GC. Interactions of macaque blastocysts with epithelial cells in vitro. Hum. Reprod. 2005 Nov;20(11):3026-32.
44. 44. Morris SA, Grewal S, Barrios F, et al. Dynamics of anterior-posterior axis formation in the developing mouse embryo. Nat. Commun. 2012 Feb 14;3:673.
45. 45. Matthews KR, Moralí D. National human embryo and embryoid research policies: a survey of 22 top research-intensive countries. Regen. Med. 2020 Jul;15(7):1905-17.
46. 46. Rivron N, Pera M, Rossant J, Martinez Arias A, Zernicka-Goetz M, Fu J, van den Brink S, Bredenoord A, Dondorp W, de Wert G, Hyun I, Munsie M, Isasi R. Debate ethics of embryo models from stem cells. Nature. 2018 Dec;564(7735):183-5.
47. 47. Hyun I, Wilkerson A, Johnston J. Embryology policy: Revisit the 14-day rule. Nature. 2016 May 12;533(7602):169-71.
48. 48. Aach J, Lunshof J, Iyer E, et al. Addressing the ethical issues raised by synthetic human entities with embryo-like features. Elife. 2017 Mar 21;6:e20674.
49. 49. Chan S. How and Why to Replace the 14-Day Rule. Curr. Stem Cell Rep. 2018;4(3):228-34.
50. 50. Hurlbut JB, Hyun I, Levine AD, et al. Revisiting the Warnock rule. Nat. Biotechnol. 2017 Nov 9;35(11):1029-42.
51. 51. Appleby JB, Bredenoord AL. Should the 14-day rule for embryo research become the 28-day rule? EMBO Mol. Med. 2018 Sep;10(9):e9437.
52. 52. Cavaliere G. A 14-day limit for bioethics: the debate over human embryo research. BMC Med. Ethics. 2017 May 30;18(1):38.
53. 53. Pera MF. Human embryo research and the 14-day rule. Development. 2017 Jun 1;144(11):1923-5.
54. 54. Williams K, Johnson MH. Adapting the 14-day rule for embryo research to encompass evolving technologies. Reprod. Biomed Soc. Online. 2020 Jan 21;10:1-9.
55. 55. Matthews KR, Rowland ML. Stem cell policy in the Obama age: UK and US perspectives. Regen. Med. 2011 Jan;6(1):125-32.
56. 56. Simunovic M, Brivanlou AH. Embryoids, organoids and gastruloids: new approaches to understanding embryogenesis. Development. 2017 Mar 15;144(6):976-85.
57. 57. Shahbazi MN, Siggia ED, Zernicka-Goetz M. Self-organization of stem cells into embryos: A window on early mammalian development. Science. 2019 Jun 7;364(6444):948-51.
58. 58. Rossant J, Tam PPL. Exploring early human embryo development. Science. 2018 Jun 8;360(6393):1075-6.
59. 59. Wallingford JB. The 200-year effort to see the embryo. Science. 2019 Aug 23;365(6455):758-9.
60. 60. Heemskerk I, Warmflash A. Pluripotent stem cells as a model for embryonic patterning: From signaling dynamics to spatial organisation in a dish. Dev. Dyn. 2016 Oct;245(10):976-90.
61. 61. Zheng Y, Xue X, Shao Y, et al. Controlled modelling of human epiblast and amnion development using stem cells. Nature. 2019 Sep;573(7774):421-5.
62. 62. Wysocka J, Rossant J. 2018 ISSCR Strategic Planning: Looking to the Future. Stem Cell Reports. 2019 Jun 11;12(6):1183-5.
63. 63. Laurent J, Blin G, Chatelain F, et al. Convergence of microengineering and cellular self-organisation towards functional tissue manufacturing. Nat. Biomed. Eng. 2017 Dec;1(12):939-56.
64. 64. Sozen B, Amadei G, Cox A, et al. Self-assembly of embryonic and two extra-embryonic stem cell types into gastrulating embryo-like structures. Nat. Cell Biol. 2018 Aug;20(8):979-89.
65. 65. Hyun I, Munsie M, Pera MF, Rivron NC, Rossant J. Toward Guidelines for Research on Human Embryo Models Formed from Stem Cells. Stem Cell Reports. 2020 Feb 11;14(2):169-74.
66. 66. Pera MF, de Wert G, Dondorp W, et al. What if stem cells turn into embryos in a dish? Nat. Methods. 2015 Oct;12(10):917-9.