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Am J Perinatol 2022; 39(11): 1212-1222
DOI: 10.1055/s-0040-1721718
Original Article

Human Mesenchymal Stem Cell Sheets Improve Uterine Incision Repair in a Rodent Hysterotomy Model

Goro Kuramoto
1   Cell Sheet Tissue Engineering Center, Department of Pharmaceutics and Pharmaceutical Chemistry, Health Sciences, University of Utah, Salt Lake City, Utah
,
Ibrahim A. Hammad 
2   Intermountain Healthcare, Salt Lake City, Utah
3   Division of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Health Sciences, University of Utah, Salt Lake City, Utah
,
Brett D. Einerson
2   Intermountain Healthcare, Salt Lake City, Utah
3   Division of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Health Sciences, University of Utah, Salt Lake City, Utah
,
Amanda A. Allshouse
3   Division of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Health Sciences, University of Utah, Salt Lake City, Utah
,
Michelle Debbink
2   Intermountain Healthcare, Salt Lake City, Utah
3   Division of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Health Sciences, University of Utah, Salt Lake City, Utah
,
David W. Grainger
1   Cell Sheet Tissue Engineering Center, Department of Pharmaceutics and Pharmaceutical Chemistry, Health Sciences, University of Utah, Salt Lake City, Utah
4   Department of Biomedical Engineering, University of Utah, Salt Lake City, Utah
,
Robert M. Silver
2   Intermountain Healthcare, Salt Lake City, Utah
3   Division of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Health Sciences, University of Utah, Salt Lake City, Utah
,
Teruo Okano
1   Cell Sheet Tissue Engineering Center, Department of Pharmaceutics and Pharmaceutical Chemistry, Health Sciences, University of Utah, Salt Lake City, Utah
5   Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan
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Abstract

Objective The study aimed to assess the feasibility of creating and transplanting human umbilical cord mesenchymal stem cell sheets applied to a rat model of hysterotomy, and additionally to determine benefits of human umbilical cord mesenchymal stem cell sheet transplantation in reducing uterine fibrosis and scarring.

Study Design Human umbilical cord mesenchymal stem cell sheets are generated by culturing human umbilical cord mesenchymal stem cells on thermo-responsive cell culture plates. The temperature-sensitive property of these culture dishes facilitates normal cell culture in a thin contiguous layer and allows for reliable recovery of intact stem cell sheets without use of destructive proteolytic enzymes.

We developed a rat hysterotomy model using nude rats. The rat uterus has two distinct horns: one horn provided a control/untreated scarring site, while the second horn was the cell sheet transplantation site.

On day 14 following surgery, complete uteri were harvested and subjected to histologic evaluations of all hysterotomy sites.

Results The stem cell sheet culture process yielded human umbilical cord mesenchymal stem cell sheets with surface area of approximately 1 cm2.

Mean myometrial thickness in the cell sheet-transplanted group was 274 μm compared with 191 μm in the control group (p = 0.02). Mean fibrotic surface area in the human umbilical cord mesenchymal stem cell sheet-transplanted group was 95,861 μm2 compared with 129,185 μm2 in the control group. Compared with control horn sites, cell sheet-transplanted horns exhibited significantly smaller fibrotic-to-normal myometrium ratios (0.18 vs. 0.27, respectively, p = 0.029). Mean number of fibroblasts in cell sheet-transplanted horns was significantly smaller than the control horns (483 vs. 716/mm2, respectively, p = 0.001).

Conclusion Human umbilical cord mesenchymal stem cell sheet transplantation is feasible in a rat model of hysterotomy. Furthermore, use of stem cell sheets reduces fibroblast infiltration and uterine scar fibrotic tissue formation during hysterotomy healing, potentially mitigating risks of uterine scar formation.

Key Points

  • Stem cell sheet transplanted to hysterotomy promotes myometrial regeneration and reduced fibrotic tissue formation.

  • This study demonstrates the feasibility of using human umbilical cord mesenchymal stem cell sheets.

Keywords

cell therapy - cell sheet therapy - scar repair - cesarean section - placenta accrete - uterine rupture - fibrosis - regenerative medicine

Note

T.O. is a founder of CellSeed Inc. which has licenses for the production of certain cell sheet-related technologies and patents from Tokyo Women's Medical University (Japan).


An earlier version of this work was presented at the 2019 Society for Maternal Fetal Medicine Annual Meeting in Las Vegas, NV from February 11 to 16, 2019 (available at: https://www.ajog.org/article/S0002-9378(18)32011-8/pdf).




Publikationsverlauf

Eingereicht: 14. März 2020

Angenommen: 04. November 2020

Artikel online veröffentlicht:
27. Dezember 2020

© 2020. Thieme. All rights reserved.

Thieme Medical Publishers, Inc.
333 Seventh Avenue, 18th Floor, New York, NY 10001, USA

 

  • References

  • 1 Hall MJ, DeFrances CJ, Williams SN, Golosinskiy A, Schwartzman A. National Hospital Discharge Survey: 2007 summary. Natl Health Stat Rep 2010; (29) 1-20 , 24
    PubMedGoogle Scholar
  • 2 Silver RM, Landon MB, Rouse DJ. et al; National Institute of Child Health and Human Development Maternal-Fetal Medicine Units Network. Maternal morbidity associated with multiple repeat cesarean deliveries. Obstet Gynecol 2006; 107 (06) 1226-1232
    Google Scholar
  • 3 Clark EA, Silver RM. Long-term maternal morbidity associated with repeat cesarean delivery. Am J Obstet Gynecol 2011; 205 (6, Suppl): S2-S10
    CrossrefPubMedGoogle Scholar
  • 4 Boerma T, Ronsmans C, Melesse DY. et al. Global epidemiology of use of and disparities in caesarean sections. Lancet 2018; 392 (10155): 1341-1348
    CrossrefPubMedGoogle Scholar
  • 5 Martin JA, Hamilton BE, Osterman MJK, Driscoll AK, Drake P. Births: final data for 2017. Natl Vital Stat Rep 2018; 67 (08) 1-50
    Google Scholar
  • 6 Wu S, Kocherginsky M, Hibbard JU. Abnormal placentation: twenty-year analysis. Am J Obstet Gynecol 2005; 192 (05) 1458-1461
    Google Scholar
  • 7 Bailit JL, Grobman WA, Rice MM. et al; Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD) Maternal-Fetal Medicine Units (MFMU) Network. Morbidly adherent placenta treatments and outcomes. Obstet Gynecol 2015; 125 (03) 683-689
    Google Scholar
  • 8 Al-Zirqi I, Stray-Pedersen B, Forsén L, Daltveit AK, Vangen S. Uterine rupture: trends over 40 years. BJOG 2016; 123 (05) 780-787
    CrossrefPubMedGoogle Scholar
  • 9 Al-Zirqi I, Daltveit AK, Vangen S. Maternal outcome after complete uterine rupture. Acta Obstet Gynecol Scand 2019; 98 (08) 1024-1031
    CrossrefPubMedGoogle Scholar
  • 10 Al-Zirqi I, Daltveit AK, Vangen S. Infant outcome after complete uterine rupture. Am J Obstet Gynecol 2018; 219 (01) 109.e1-109.e8
    CrossrefPubMedGoogle Scholar
  • 11 Joseph KS, Young DC, Dodds L. et al. Changes in maternal characteristics and obstetric practice and recent increases in primary cesarean delivery. Obstet Gynecol 2003; 102 (04) 791-800
    PubMedGoogle Scholar
  • 12 Habiba M, Kaminski M, Da Frè M. et al. Caesarean section on request: a comparison of obstetricians' attitudes in eight European countries. BJOG 2006; 113 (06) 647-656
    CrossrefPubMedGoogle Scholar
  • 13 Murthy K, Grobman WA, Lee TA, Holl JL. Association between rising professional liability insurance premiums and primary cesarean delivery rates. Obstet Gynecol 2007; 110 (06) 1264-1269
    CrossrefPubMedGoogle Scholar
  • 14 Hauth JC, Owen J, Davis RO. Transverse uterine incision closure: one versus two layers. Am J Obstet Gynecol 1992; 167 (4 Pt 1): 1108-1111
    CrossrefPubMedGoogle Scholar
  • 15 Harrigill KM, Miller HS, Haynes DE. The effect of intraabdominal irrigation at cesarean delivery on maternal morbidity: a randomized trial. Obstet Gynecol 2003; 101 (01) 80-85
    PubMedGoogle Scholar
  • 16 Walfisch A, Beloosesky R, Shrim A, Hallak M. Adhesion prevention after cesarean delivery: evidence, and lack of it. Am J Obstet Gynecol 2014; 211 (05) 446-452
    CrossrefPubMedGoogle Scholar
  • 17 Jauniaux E, Jurkovic D. Placenta accreta: pathogenesis of a 20th century iatrogenic uterine disease. Placenta 2012; 33 (04) 244-251
    CrossrefPubMedGoogle Scholar
  • 18 Ho CH, Lan CW, Liao CY, Hung SC, Li HY, Sung YJ. Mesenchymal stem cells and their conditioned medium can enhance the repair of uterine defects in a rat model. J Chin Med Assoc 2018; 81 (03) 268-276
    CrossrefPubMedGoogle Scholar
  • 19 Kim K, Bou-Ghannam S, Thorp H, Grainger DW, Okano T. Human mesenchymal stem cell sheets in xeno-free media for possible allogenic applications. Sci Rep 2019; 9 (01) 14415
    CrossrefPubMedGoogle Scholar
  • 20 Nakao M, Kim K, Nagase K, Grainger DW, Kanazawa H, Okano T. Phenotypic traits of mesenchymal stem cell sheets fabricated by temperature-responsive cell culture plate: structural characteristics of MSC sheets. Stem Cell Res Ther 2019; 10 (01) 353
    CrossrefPubMedGoogle Scholar
  • 21 Parekkadan B, Milwid JM. Mesenchymal stem cells as therapeutics. Annu Rev Biomed Eng 2010; 12: 87-117
    CrossrefPubMedGoogle Scholar
  • 22 Ra JC, Shin IS, Kim SH. et al. Safety of intravenous infusion of human adipose tissue-derived mesenchymal stem cells in animals and humans. Stem Cells Dev 2011; 20 (08) 1297-1308
    CrossrefPubMedGoogle Scholar
  • 23 Kurtz A. Mesenchymal stem cell delivery routes and fate. Int J Stem Cells 2008; 1 (01) 1-7
    CrossrefPubMedGoogle Scholar
  • 24 Braid LR, Wood CA, Wiese DM, Ford BN. Intramuscular administration potentiates extended dwell time of mesenchymal stromal cells compared to other routes. Cytotherapy 2018; 20 (02) 232-244
    CrossrefPubMedGoogle Scholar
  • 25 Devine SM, Cobbs C, Jennings M, Bartholomew A, Hoffman R. Mesenchymal stem cells distribute to a wide range of tissues following systemic infusion into nonhuman primates. Blood 2003; 101 (08) 2999-3001
    CrossrefPubMedGoogle Scholar
  • 26 Reinecke H, Murry CE. Taking the death toll after cardiomyocyte grafting: a reminder of the importance of quantitative biology. J Mol Cell Cardiol 2002; 34 (03) 251-253
    CrossrefPubMedGoogle Scholar
  • 27 Terrovitis JV, Smith RR, Marbán E. Assessment and optimization of cell engraftment after transplantation into the heart. Circ Res 2010; 106 (03) 479-494
    CrossrefPubMedGoogle Scholar
  • 28 Elman JS, Murray RC, Wang F. et al. Pharmacokinetics of natural and engineered secreted factors delivered by mesenchymal stromal cells. PLoS One 2014; 9 (02) e89882
    CrossrefPubMedGoogle Scholar
  • 29 Shimizu T. Cell sheet-based tissue engineering for fabricating 3-dimensional heart tissues. Circ J 2014; 78 (11) 2594-2603
    CrossrefPubMedGoogle Scholar
  • 30 Kanai N, Yamato M, Ohki T, Yamamoto M, Okano T. Fabricated autologous epidermal cell sheets for the prevention of esophageal stricture after circumferential ESD in a porcine model. Gastrointest Endosc 2012; 76 (04) 873-881
    CrossrefPubMedGoogle Scholar
  • 31 Takahashi H, Okano T. Thermally-triggered fabrication of cell sheets for tissue engineering and regenerative medicine. Adv Drug Deliv Rev 2019; 138: 276-292
    CrossrefPubMedGoogle Scholar
  • 32 Matsuura K, Haraguchi Y, Shimizu T, Okano T. Cell sheet transplantation for heart tissue repair. J Control Release 2013; 169 (03) 336-340
    CrossrefPubMedGoogle Scholar
  • 33 Miyahara Y, Nagaya N, Kataoka M. et al. Monolayered mesenchymal stem cells repair scarred myocardium after myocardial infarction. Nat Med 2006; 12 (04) 459-465
    CrossrefPubMedGoogle Scholar
  • 34 Hida N, Nishiyama N, Miyoshi S. et al. Novel cardiac precursor-like cells from human menstrual blood-derived mesenchymal cells. Stem Cells 2008; 26 (07) 1695-1704
    CrossrefPubMedGoogle Scholar
  • 35 Kaibuchi N, Iwata T, Yamato M, Okano T, Ando T. Multipotent mesenchymal stromal cell sheet therapy for bisphosphonate-related osteonecrosis of the jaw in a rat model. Acta Biomater 2016; 42: 400-410
    CrossrefPubMedGoogle Scholar
  • 36 Kawamura M, Miyagawa S, Fukushima S. et al. Xenotransplantation of bone marrow-derived human mesenchymal stem cell sheets attenuates left ventricular remodeling in a porcine ischemic cardiomyopathy model. Tissue Eng Part A 2015; 21 (15-16): 2272-2280
    CrossrefPubMedGoogle Scholar
  • 37 Kong P, Christia P, Saxena A, Su Y, Frangogiannis NG. Lack of specificity of fibroblast-specific protein 1 in cardiac remodeling and fibrosis. Am J Physiol Heart Circ Physiol 2013; 305 (09) H1363-H1372
    CrossrefPubMedGoogle Scholar
  • 38 Österreicher CH, Penz-Österreicher M, Grivennikov SI. et al. Fibroblast-specific protein 1 identifies an inflammatory subpopulation of macrophages in the liver. Proc Natl Acad Sci U S A 2011; 108 (01) 308-313
    CrossrefPubMedGoogle Scholar
  • 39 Patel AN, Vargas V, Revello P, Bull DA. Mesenchymal stem cell population isolated from the subepithelial layer of umbilical cord tissue. Cell Transplant 2013; 22 (03) 513-519
    CrossrefPubMedGoogle Scholar
  • 40 Tuma J, Carrasco A, Castillo J. et al. RESCUE-HF trial: retrograde delivery of allogeneic umbilical cord lining subepithelial cells in patients with heart failure. Cell Transplant 2016; 25 (09) 1713-1721
    CrossrefPubMedGoogle Scholar
  • 41 Bartolucci J, Verdugo FJ, González PL. et al. Safety and efficacy of the intravenous infusion of umbilical cord mesenchymal stem cells in patients with heart failure: a phase 1/2 randomized controlled trial (RIMECARD trial [randomized clinical trial of intravenous infusion umbilical cord mesenchymal stem cells on cardiopathy]). Circ Res 2017; 121 (10) 1192-1204
    CrossrefPubMedGoogle Scholar
  • 42 La Rocca G, Anzalone R, Corrao S. et al. Isolation and characterization of Oct-4+/HLA-G+ mesenchymal stem cells from human umbilical cord matrix: differentiation potential and detection of new markers. Histochem Cell Biol 2009; 131 (02) 267-282
    CrossrefPubMedGoogle Scholar
  • 43 Ishitani A, Sageshima N, Hatake K. The involvement of HLA-E and -F in pregnancy. J Reprod Immunol 2006; 69 (02) 101-113
    CrossrefPubMedGoogle Scholar
  • 44 Matsuura K, Utoh R, Nagase K, Okano T. Cell sheet approach for tissue engineering and regenerative medicine. J Control Release 2014; 190: 228-239
    CrossrefPubMedGoogle Scholar
  • 45 Sekine H, Shimizu T, Dobashi I. et al. Cardiac cell sheet transplantation improves damaged heart function via superior cell survival in comparison with dissociated cell injection. Tissue Eng Part A 2011; 17 (23-24): 2973-2980
    CrossrefPubMedGoogle Scholar
  • 46 Narita T, Shintani Y, Ikebe C. et al. The use of scaffold-free cell sheet technique to refine mesenchymal stromal cell-based therapy for heart failure. Mol Ther 2013; 21 (04) 860-867
    CrossrefPubMedGoogle Scholar
  • 47 Fukushima T, Uchiyama S, Tanaka H, Kataoka H. Hepatocyte growth factor activator: a proteinase linking tissue injury with repair. Int J Mol Sci 2018; 19 (11) E3435
    CrossrefPubMedGoogle Scholar
  • 48 Wilgus TA. Vascular endothelial growth factor and cutaneous scarring. Adv Wound Care (New Rochelle) 2019; 8 (12) 671-678
    CrossrefPubMedGoogle Scholar
  • 49 Kim SR, Yi HJ, Lee YN. et al. Engineered mesenchymal stem-cell-sheets patches prevents postoperative pancreatic leakage in a rat model. Sci Rep 2018; 8 (01) 360
    CrossrefPubMedGoogle Scholar
  • 50 Kato Y, Iwata T, Washio K. et al. Creation and transplantation of an adipose-derived stem cell (ASC) sheet in a diabetic wound-healing model, J Vis Exp. 2017 54539. 126
    PubMedGoogle Scholar
  • 51 Kim K, Thorp H, Bou-Ghannam S, Grainger DW, Okano T. Stable cell adhesion affects mesenchymal stem cell sheet fabrication: effects of fetal bovine serum and human platelet lysate. J Tissue Eng Regen Med 2020; 14 (05) 741-753
    CrossrefPubMedGoogle Scholar
  • 52 Lee JK, Jin HK, Endo S, Schuchman EH, Carter JE, Bae JS. Intracerebral transplantation of bone marrow-derived mesenchymal stem cells reduces amyloid-beta deposition and rescues memory deficits in Alzheimer's disease mice by modulation of immune responses. Stem Cells 2010; 28 (02) 329-343
    CrossrefPubMedGoogle Scholar
  • 53 Djouad F, Bouffi C, Ghannam S, Noël D, Jorgensen C. Mesenchymal stem cells: innovative therapeutic tools for rheumatic diseases. Nat Rev Rheumatol 2009; 5 (07) 392-399
    CrossrefPubMedGoogle Scholar
  • 54 Bowers D, McKenzie D, Dutta D, Wheeless CR, Cohen WR. Growth hormone treatment after cesarean delivery in rats increases the strength of the uterine scar. Am J Obstet Gynecol 2001; 185 (03) 614-617
    CrossrefPubMedGoogle Scholar