Austin J In Vitro Fertili. 2016; 3(1): 1025.
Isolation, Culture and Characterization of Human Endometrial Mesenchymal Stem/ Stromal Cells (EnMSCs): A Mini Review
Akyash F¹, Sadeghian-Nodoushan F¹ and Aflatoonian B1,2*
¹Stem Cell Biology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
²Department of Advanced Medical Sciences and Technologies, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
*Corresponding author: Aflatoonian B, Stem Cell Biology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Received: February 25, 2016; Accepted: April 23, 2016; Published: April 25, 2016
The endometrium is an extraordinary tissue with the dynamic remodeling capacity in each menstrual cycle for the purpose of embryo implantation. These preparing undergone consecutive changes of sex hormones during monthly periods. Endometrial basal layer cells could proliferate and differentiate into various cell types. This ability of endometrium confirmed the appearance of specific population of cells could regenerate functional is layer, known as, stem cells. The progression of infertility treatments with novel therapies for the patient with endometrial dysfunction requires an appropriate understanding of the role of endometrial stem cells in regenerative medicine and subsequent cell therapy.
Keywords: Cell Therapy; Endometrial Stem/Stromal Cells; Regenerative Medicine; Uterus
ASCs: Adult Stem Cell; EnMSCs: Endometrial Mesenchymal Stem/ Stromal Cells; CFU: Colony Forming Unit; MSCs: Mesenchymal Stem Cell; MS: Multiple Sclerosis
Adult Stem Cells (ASCs) have functional properties including: self-renewal, proliferation and differentiation capacity into one or more lineage [1,2]. These cells could be isolate from different tissue origins such as: intestines , muscles , skin , blood , nervous system [7-9], and endometrium .
The human endometrium is a unique model with considerable regenerative potential during women’s reproductive life. The endometrium undergoes dynamic remodeling including regeneration, differentiation, tissue break down and shedding of upper functional in different phases of each menstrual cycle. As we all aware, regeneration of the new functional is part for subsequent cycle could performed with remaining germinal portion in the lower basalis layer . Consequently, It has been hypothesized basal layer of human endometrium remaining to regeneration potential contain epithelial and stromal adult stem cells, which after shedding of top two-third at menstruation, these cells population facilitated regeneration of endometrial glandular tissue which protected by vascularized stroma [12,13]. Therefore, in this mini-review we briefly focused on the identification and characterization of Endometrial Mesenchymal Stem/Stromal Cells (EnMSCs) population according current studies and use their regeneration potential in future medical therapeutics.
Resembling to other ASCs, EnMSCs have Colony Forming Unit (CFU) activity, which could determine adult stem cell characteristics like: self-renewal, differentiation, and high proliferative potency in vitro . This confirmed their ability to response to the monthly steroid hormonal changes and differentiation the endometrium for fertilized egg receptivity. Regeneration process after shedding of the functionalis layer during each menstrual cycle, emphasized to reside of epithelial and stromal CFU in the basalis layer . Another study reported that post-menopausal endometrium, whether atrophic with low level of estrogen circulation, or after estrogen treatment, contains a small population of stromal cells with Mesenchymal Stem Cell (MSCs) characteristics .
Cloning assays shows the efficiency of each potential stem cell marker to enrich for endometrial stromal CFU. CD146 could be as a potential marker enriching for endometrial stromal CFU, and detection of CD90hi versus CD90lo stromal cells in all samples may be of value for further investigation as a potential marker . The identification of STRO-1 and CD133 as negative markers of endometrial stromal CFU validates the importance of undertaking functional stem cell assays to demonstrate the utility of the stem cell markers for isolation of purified stem cell populations. The same study shows which endometrial stromal colony-forming cells were identified in perivascular space, even near the microvasculature . Isolation of EnMSCs from human endometrial tissue after in vitro culture showed expression of MSCs markers CD90, CD105 but hematopoietic markers like CD34. CD45 did not express . Another group reported same results. In this study isolated cells after flow cytometry analysis showed positive MSCs markers (CD146, CD90 and CD105), but hematopoietic and endothelial markers CD34 and CD31 respectively, were negative [18,19] (Figure 1).
Figure 1: CFU assay (A), culture (B) and characterization (C) of human EnMSCs.
Regenerative medicine is a hopeful policy for human cells, tissues, organs replacement or regeneration and finally reestablishment of normal function [20,21]. To prevention the limitation of in vivo study, endometrium generally has been studied in vitro with cell culture and tissue engineering techniques to demonstrate physiological and disorders of the endometrium . For evaluation of regenerative potential of EnMSCs in Immunodeficient mice with Duchenne muscular dystrophy, these cells after transplantation into atrophied skeletal muscle fibers, contributed to muscle repair . However, exact mechanism of repair has not been clear, it was claimed that fusion and in situ differentiation of cells might have been involved, and because of transplanted EnMSCs could stimulated angiogenesis in peri-muscle fibers regions . Furthermore, in patient that suffering from Duchenne muscular dystrophy, EnMSCs injected into muscles and reported improvement in muscle strength and reduction of respiratory infections . Other report supported the role of EnMSCs in angiogenesis procedure, which showed these cells improved limb ischemia by femoral artery ligation . Hida and co-workers used EnMSCs for treatment of murine with myocardial infarction . In this study, comparison between treated infarct area with EnMSCs and bone marrow MSCs was evaluated and results showed, grafted EnMSCs into the infarct area differentiated into a-actinin+, troponin+ striated cardiac muscle cells and more improvement in this area was observed rather than control bone marrow MSCs group . Another study that used EnMSCs for tissue engineering, reported reconstruction of women urinary bladder wall with constructed nano fibrous silk-collagen fibers and cultured EnMSCs, these cells could differentiate into smooth muscle cells  Zhong et al, demonstrated the application of EnMSCs that derived from menstrual bleeding and role of these cells in reduction of immunological reactions after intravenous and intrathechal injection for patient with Multiple Sclerosis (MS) disease .
Endometrial tissue is a part of uterus that undergoes a cyclical regeneration every month in normal women’s life span. Endometrial regeneration after delivery even in postmenopausal women after estrogen therapy indicates that EnMSCs could proliferate and differentiate into different cell types and confirmed the stemness potential of these cells. Hence, based on current studies, EnMSCs can be used in the future for novel therapeutic methods in regenerative medicine such as treatment of uterine-factor infertile female that suffering from infertility.
- Eckfeldt CE, Mendenhall EM, Verfaillie CM. The molecular repertoire of the 'almighty' stem cell. Nat Rev Mol Cell Biol. 2005; 6: 726-737.
- Shostak S. (Re)defining stem cells. Bioessays. 2006; 28: 301-308.
- Bjerknes M, Cheng H. Clonal analysis of mouse intestinal epithelial progenitors. Gastroenterology. 1999; 116: 7-14.
- Jankowski RJ, Deasy BM, Huard J. Muscle-derived stem cells. Gene Ther. 2002; 9: 642-647.
- Alonso L, Fuchs E. Stem cells of the skin epithelium. Proc Natl Acad Sci USA. 2003; 100: 11830-11835.
- Spangrude GJ, Smith L, Uchida N, Ikuta K, Heimfeld S, Friedman J, et al. Mouse hematopoietic stem cells. Blood. 1991; 78: 1395-1402.
- Morrison SJ, White PM, Zock C, Anderson DJ. Prospective identification, isolation by flow cytometry, and in vivo self-renewal of multipotent mammalian neural crest stem cells. Cell. 1999; 96: 737-749.
- Uchida N, Buck DW, He D, Reitsma MJ, Masek M, Phan TV, et al. Direct isolation of human central nervous system stem cells. Proc Natl Acad Sci USA. 2000; 97: 14720-14725.
- Rietze RL, Valcanis H, Brooker GF, Thomas T, Voss AK, Bartlett PF. Purification of a pluripotent neural stem cell from the adult mouse brain. Nature. 2001; 412: 736-739.
- Chan RW, Gargett CE. Identification of label-retaining cells in mouse endometrium. Stem Cells. 2006; 24: 1529-1538.
- Padykula HA, Coles LG, McCracken JA, King NW Jr, Longcope C, Kaiserman-Abramof IR. A zonal pattern of cell proliferation and differentiation in the rhesus endometrium during the estrogen surge. Biol Reprod. 1984; 31: 1103-1118.
- Padykula HA. Regeneration in the primate uterus: the role of stem cells. Ann N Y Acad Sci. 1991; 622: 47-56.
- Gargett CE. Uterine stem cells: what is the evidence? Hum Reprod Update. 2007; 13: 87-101.
- Gargett CE, Schwab KE, Zillwood RM, Nguyen HP, Wu D. Isolation and culture of epithelial progenitors and mesenchymal stem cells from human endometrium. Biol Reprod. 2009; 80: 1136-1145.
- Ulrich D, Tan KS, Deane J, Schwab K, Cheong A, Rosamilia A, et al. Mesenchymal stem/stromal cells in post-menopausal endometrium. Hum Reprod. 2014; 29: 1895-1905.
- Schwab KE, Hutchinson P, Gargett CE. Identification of surface markers for prospective isolation of human endometrial stromal colony-forming cells. Hum Reprod. 2008; 23: 934-943.
- Wang L, Yang X, Wang W. [Isolation and identification of human endometrial stromal stem cells]. Zhejiang Da Xue Xue Bao Yi Xue Ban. 2013; 42: 311-318.
- Ai J, Shahverdi AR, Barough SE, Kouchesfehani HM, Heidari S, Roozafzoon R, et al. Derivation of Adipocytes from Human Endometrial Stem Cells (EnSCs). J Reprod Infertil. 2012; 13: 151-157.
- Akyash F, Aflatoonian A, Rezazadeh Valojerdi M, SadeghianNodoushan F, Aflatoonian B. In Vitro isolation, culture and identification of human endometrial mesenchymal stem/stromal cells (EnMSCs). Celljournal. 2015; 17; 22.
- Orlando G, Baptista P, Birchall M, De Coppi P, Farney A, Guimaraes-Souza NK, et al. Regenerative medicine as applied to solid organ transplantation: current status and future challenges. Transplant Int. 2011; 24: 223–232.
- Orlando G, Wood KJ, Stratta RJ, Yoo JJ, Atala A, Soker S. Regenerative medicine and organ transplantation: past, present, and future. Transplantation. 2011; 91: 1310-1317.
- Baghaban Eslami Nejad MR, Rezazadeh Valojerdi M, Kazemi Ashtiani S. A comparison of polarized and non-polarized human endometrial monolayer culture systems on murine embryo development. J Exp Clin Assist Reprod. 2005; 2: 7.
- Cui CH, Uyama T, Miyado K, Terai M, Kyo S, Kiyono T, et al. Menstrual blood-derived cells confer human dystrophin expression in the murine model of Duchenne muscular dystrophy via cell fusion and myogenic transdifferentiation. Mol Biol Cell. 2007; 18: 1586-1594.
- Ichim TE, Alexandrescu DT, Solano F, Lara F, Campion Rde N, Paris E, et al. Mesenchymal stem cells as anti-inflammatories: implications for treatment of Duchenne muscular dystrophy. Cell Immunol. 2010; 260: 75-82.
- Murphy MP, Wang H, Patel AN, Kambhampati S, Angle N, Chan K, et al. Allogeneic endometrial regenerative cells: an "Off the shelf solution" for critical limb ischemia? J Transl Med. 2008; 6: 45.
- Hida N, Nishiyama N, Miyoshi S, Kira S, Segawa K, Uyama T, et al. Novel cardiac precursor-like cells from human menstrual blood-derived mesenchymal cells. Stem Cells. 2008; 26: 1695-1704.
- Shoae-Hassani A, Sharif S, Seifalian AM, Mortazavi-Tabatabaei SA, Rezaie S, Verdi J. Endometrial stem cell differentiation into smooth muscle cell: a novel approach for bladder tissue engineering in women. BJU Int. 2013; 112: 854-863.
- Zhong Z, Patel AN, Ichim TE, Riordan NH, Wang H, Min WP, et al. Feasibility investigation of allogeneic endometrial regenerative cells. J Transl Med. 2009; 7: 15.
Citation: Akyash F, Sadeghian-Nodoushan F and Aflatoonian B. Isolation, Culture and Characterization of Human Endometrial Mesenchymal Stem/ Stromal Cells (EnMSCs): A Mini Review. Austin J In Vitro Fertili. 2016; 3(1): 1025. ISSN:2471-0628