Estandarización del aislamiento y caracterización de células estromales de decidua y útero murinos

Authors

  • Lina Cadavid
  • Cristina Ávila
  • Angela Patricia Cadavid

DOI:

https://doi.org/10.17533/udea.rccp.324497

Keywords:

BeWo cells, decidual macrophages, HeLa cells, murine decidua, stromal cells

Abstract

Summary

Stromal cells are the most abundant cell population present in decidual tissue; they are involved in key processes during embryo implantation, fetal nutrition and the pregnancy maintenance. Described procedures for stromal cells isolation require the use of many monoclonal antibodies due to contamination with another cell types in the decidua; besides, some markers of stromal cells show variability during the days of gestation. In this study, we standardized a procedure for isolation by enzymatic digestion, density gradient and adherence to plastic. Murine stromal cells were characterized by exclusion of markers that are expressed in macrophages (F4-80), epithelial cells and trophoblast (cytokeratin-7), yielding a 98% of negative cells for these markers that correspond to stromal cells. This isolation procedure permits to obtain stromal cells with less expensive and high efficiency methods that provide a useful cellular model to study the physiology of gestation in different species.

 

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References

Arnold JT, Kaufman DG, Seppala M, Lessey BA. Endometrial stromal cells regulate epithelial cell growth in vitro: a new co-culture model. Hum Reprod 16 (5): 836-45, 2001. DOI: https://doi.org/10.1093/humrep/16.5.836

Bany BM, Kennedy TG. Role of interleukin 1 in the regulation of cyclooxygenase gene expression in rat endometrial stromal cells. J Reprod Fertil 1999; 115: 125-31. DOI: https://doi.org/10.1530/jrf.0.1150125

Bowen JA, Newton GR, Weise DW, Bazer FW, Burghardt RC. Characterization of a polarized porcine uterine epithelial model system. Biol Reprod 1996; 55: 613-9. DOI: https://doi.org/10.1095/biolreprod55.3.613

Chou CS, Tai CJ, MacCalman CD, Leung PC. Dose-dependent effects of gonadotropin releasing hormone on matrix metalloproteinase (MMP)-2, and MMP-9 and tissue specifi c inhibitor of metalloproteinases-1 messenger ribonucleic acid levels in human decidual stromal cells in vitro. J Clin Endocrinol Metab 2003; 88: 680-8. DOI: https://doi.org/10.1210/jc.2002-021277

Corver WE, Ter Haar NT, Dreef EJ, Miranda NF, Prins FA, et al. High-resolution multi-parameter DNA fl ow cytometry enables detection of tumour and stromal cell subpopulations in paraffi n-embedded tissues. J Pathol 2005; 206: 233-41. DOI: https://doi.org/10.1002/path.1765

Dunn CL, Critchley HO, Kelly RW. IL-15 regulation in human endometrial stromal cells. J Clin Endocrinol Metab 2002; 87: 1898-901. DOI: https://doi.org/10.1210/jcem.87.4.8539

Fazleabas AT, Strakova Z. Endometrial function: cell specifi c changes in the uterine environment. Mol Cell Endocrinol 2002; 186: 143-7. DOI: https://doi.org/10.1016/S0303-7207(01)00655-4

Geggel HS, Gipson IK. Removal of viable sheets of conjunctival epithelium with dispase II. Invest Ophthalmol Vis Sci 1985; 26: 15-22.

Hwang JH, Park MI, Hwang YY, Yoo HJ, Mardon HJ. The characteristics of integrins expression in decidualized human endometrial stromal cell induced by 8-Br-cAMP in in vitro. Exp Mol Med 2002; 34: 194-200. DOI: https://doi.org/10.1038/emm.2002.28

Kawano Y, Nakamura S, Nasu K, Narahara H, Miyakawa I. The effect of epidermal growth factor on vascular endothelial growth factor secretion by endometrial stromal cells. Clin Exp Med 2002; 2: 69-75. DOI: https://doi.org/10.1007/s102380200009

King A. Uterine leukocytes and decidualization. Hum Reprod Update 2000; 6: 28-36, DOI: https://doi.org/10.1093/humupd/6.1.28

.Komatsu T, Konishi I, Mandai M, Mori T, Hiai H, Fukumoto M. Expression of class I human leukocyte antigen (HLA) and beta2-microglobulin is associated with decidualization of human endometrial stromal cells. Hum Reprod 1998; 13: 2246-51. DOI: https://doi.org/10.1093/humrep/13.8.2246

Nasu K, Matsui N, Narahara H, Tanaka Y, Miyakawa I. Effects of interferon-gamma on cytokine production by endometrial stromal cells. Hum Reprod 1998; 13: 2598-601. DOI: https://doi.org/10.1093/humrep/13.9.2598

Nasu K, Narahara H, Matsui N, Kawano Y, Tanaka Y, Miyakawa I. Platelet-activating factor stimulates cytokine production by human endometrial stromal cells. Mol Hum Reprod 1999; 5: 548-53. DOI: https://doi.org/10.1093/molehr/5.6.548

Nasu K, Sugano T, Fujisawa K, Arima K, Narahara H, Miyakawa I. Effects of interleukin-4 on the in-vitro production of cytokines by human endometrial stromal cells. Mol Hum Reprod 2001; 7: 265-70. DOI: https://doi.org/10.1093/molehr/7.3.265

Nasu K, Arima K, Fujisawa K, Nishida M, Kai K, Miyakawa I. Secretion of keratinocyte growth factor by cultured human endometrial stromal cells is induced through a cyclic adenosine monophosphate-dependent pathway. Fertil Steril 2002; 77: 392-5. DOI: https://doi.org/10.1016/S0015-0282(01)02975-2

Nasu K, Fukuda J, Sun B, Nishida M, Miyakawa I. Interleukin-13 and tumor necrosis factor-beta differentially regulate the production of cytokines by cultured human endometrial stromal cells. Fertil Steril 2003; 79: 821-7. DOI: https://doi.org/10.1016/S0015-0282(02)04831-8

Oliver C, Cowdrey N, Abadia-Molina AC, Olivares EG. Antigen phenotype of cultured decidual stromal cells of human term decidua. J Reprod Immunol 1999; 45 (1): 19-30. DOI: https://doi.org/10.1016/S0165-0378(99)00041-8

Vigano P, Somigliana E, Mangioni S, Vignali M, Di Blasio AM. Expression of interleukin-10 and its receptor is up-regulated in early pregnant versus cycling human endometrium. J Clin Endocrinol Metab 2002; 87: 5730-6. DOI: https://doi.org/10.1210/jc.2002-020435

Yoshino O, Osuga Y, Hirota Y, Koga K, Hirata T, et al. Endometrial stromal cells undergoing decidualization down-regulate their properties to produce proinfl ammatory cytokines in response to interleukin-1 beta via reduced p38 mitogen-activated protein kinase phosphorylation. J Clin Endocrinol Metab 2003; 88: 2236-41. DOI: https://doi.org/10.1210/jc.2002-021788

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Published

2009-12-21

How to Cite

Cadavid, L., Ávila, C., & Cadavid, A. P. (2009). Estandarización del aislamiento y caracterización de células estromales de decidua y útero murinos. Revista Colombiana De Ciencias Pecuarias, 22(4), 10. https://doi.org/10.17533/udea.rccp.324497

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Original research articles