An Human Placental Vessel Perfusion Method to Study Mesenchymal Stem/stromal Cell Migration

Overview

Journal Stem Cell Investig

Publisher AME Publishing Company

Date 2019 Mar 8

PMID 30842969

Citations 1

Authors

Balta Al-Sowayan

Rosemary J Keogh

Mohammed Abumaree

Harry M Georgiou

Bill Kalionis

Affiliations

Soon will be listed here.

Abstract

Background: To initiate tissue repair, mesenchymal stem/stromal cells (MSCs) must enter the blood stream, migrate to the targeted area, cross the endothelial barrier and home to the damaged tissue. This process is not yet fully understood in humans and thus, the aim of this study was to develop an placental vessel perfusion method to examine human MSC movement from a blood vessel into human tissue. This will provide a better understanding of MSC migration, movement through the endothelial barrier and engraftment into target tissue, in a setting that more closely represents the state, compared with conventional human cell culture models. Moreover, important similarities and differences to animal experimental model systems may be revealed by this method.

Methods: Human placental hTERT transformed MSC lines were labelled with live-cell fluorescence dyes, and then perfused into term human placental blood vessel. After labelled MSCs were perfused into the vessel, the vessel was dissected from the placenta and incubated at cell growth conditions. Following incubation, the vessel was washed thoroughly to remove unattached, labelled MSCs and then snap frozen for sectioning. After sectioning, immunofluorescence staining of the endothelium was carried out to detect if labelled MSCs crossed the endothelial barrier.

Results: Twelve placental vessel perfusions were successfully completed. In eight of the twelve perfused vessels, qualitative assessment of immunofluorescence in sections (n=20, 5 µm sections/vessel) revealed labelled MSCs had crossed the endothelial barrier.

Conclusions: The human placental vessel perfusion method could be used to assess human MSC migration into human tissue. Cells of the MSC lines were able to adhere and transmigrate through the endothelial barrier in a manner similar to that of leukocytes. Notably, cells that transmigrated remained in close proximity to the endothelium, which is consistent with the reported MSC vascular niche in placental blood vessels.

Citing Articles

Microchimerism and pregnancy complications with placental dysfunction.

Jacobsen D, Fjeldstad H, Olsen M, Sugulle M, Staff A Semin Immunopathol. 2025; 47(1):21.

PMID: 40067448 PMC: 11897092. DOI: 10.1007/s00281-025-01045-w.

Intra-Arterial Stem Cell Transplantation in Experimental Stroke in Rats: Real-Time MR Visualization of Transplanted Cells Starting With Their First Pass Through the Brain With Regard to the Therapeutic Action.

Namestnikova D, Gubskiy I, Revkova V, Sukhinich K, Melnikov P, Gabashvili A Front Neurosci. 2021; 15:641970.

PMID: 33737862 PMC: 7960930. DOI: 10.3389/fnins.2021.641970.

References

Osmond D, King R, Brennecke S, Gude N . Placental glucose transport and utilisation is altered at term in insulin-treated, gestational-diabetic patients. Diabetologia. 2001; 44(9):1133-9. DOI: 10.1007/s001250100609. View

Quintavalla J, Uziel-Fusi S, Yin J, Boehnlein E, Pastor G, Blancuzzi V . Fluorescently labeled mesenchymal stem cells (MSCs) maintain multilineage potential and can be detected following implantation into articular cartilage defects. Biomaterials. 2002; 23(1):109-19. DOI: 10.1016/s0142-9612(01)00086-2. View

Schmidt A, Ladage D, Steingen C, Brixius K, Schinkothe T, Klinz F . Mesenchymal stem cells transmigrate over the endothelial barrier. Eur J Cell Biol. 2006; 85(11):1179-88. DOI: 10.1016/j.ejcb.2006.05.015. View

Ruster B, Gottig S, Ludwig R, Bistrian R, Muller S, Seifried E . Mesenchymal stem cells display coordinated rolling and adhesion behavior on endothelial cells. Blood. 2006; 108(12):3938-44. DOI: 10.1182/blood-2006-05-025098. View

Phillipson M, Heit B, Colarusso P, Liu L, Ballantyne C, Kubes P . Intraluminal crawling of neutrophils to emigration sites: a molecularly distinct process from adhesion in the recruitment cascade. J Exp Med. 2006; 203(12):2569-75. PMC: 2118150. DOI: 10.1084/jem.20060925. View

Ley K, Laudanna C, Cybulsky M, Nourshargh S . Getting to the site of inflammation: the leukocyte adhesion cascade updated. Nat Rev Immunol. 2007; 7(9):678-89. DOI: 10.1038/nri2156. View

Steingen C, Brenig F, Baumgartner L, Schmidt J, Schmidt A, Bloch W . Characterization of key mechanisms in transmigration and invasion of mesenchymal stem cells. J Mol Cell Cardiol. 2008; 44(6):1072-1084. DOI: 10.1016/j.yjmcc.2008.03.010. View

Heinzelmann J, Enke U, Seyfarth L, Schleussner E, Malek A, Markert U . Development of a human model to study homing behavior of immune cells into decidua and placental villi under ex vivo conditions. Am J Reprod Immunol. 2008; 61(1):19-25. DOI: 10.1111/j.1600-0897.2008.00666.x. View

Barreiro O, Sanchez-Madrid F . Molecular basis of leukocyte-endothelium interactions during the inflammatory response. Rev Esp Cardiol. 2009; 62(5):552-62. DOI: 10.1016/s1885-5857(09)71837-7. View

10.

Earhart A, Patrikeeva S, Wang X, Abdelrahman D, Hankins G, Ahmed M . Transplacental transfer and metabolism of bupropion. J Matern Fetal Neonatal Med. 2009; 23(5):409-16. PMC: 2873069. DOI: 10.1080/14767050903168424. View

11.

Gahmberg C, Nortamo P, Kantor C, Autero M, Kotovuori P, Hemio L . The pivotal role of the Leu-CAM and ICAM molecules in human leukocyte adhesion. Cell Differ Dev. 1990; 32(3):239-45. DOI: 10.1016/0922-3371(90)90036-v. View

12.

May K, Grube M, Malhotra I, Long C, Singh S, Mandaliya K . Antibody-dependent transplacental transfer of malaria blood-stage antigen using a human ex vivo placental perfusion model. PLoS One. 2009; 4(11):e7986. PMC: 2777305. DOI: 10.1371/journal.pone.0007986. View

13.

Castrechini N, Murthi P, Gude N, Erwich J, Gronthos S, Zannettino A . Mesenchymal stem cells in human placental chorionic villi reside in a vascular Niche. Placenta. 2010; 31(3):203-12. DOI: 10.1016/j.placenta.2009.12.006. View

14.

Berveiller P, Mir O, Vinot C, Bonati C, Duchene P, Giraud C . Transplacental transfer of oseltamivir and its metabolite using the human perfused placental cotyledon model. Am J Obstet Gynecol. 2011; 206(1):92.e1-6. DOI: 10.1016/j.ajog.2011.07.023. View

15.

Nazarov I, Lee J, Soupene E, Etemad S, Knapik D, Green W . Multipotent stromal stem cells from human placenta demonstrate high therapeutic potential. Stem Cells Transl Med. 2012; 1(5):359-72. PMC: 3635148. DOI: 10.5966/sctm.2011-0021. View

16.

Schamberger S, Weber M, Backsch C, Sonnemann J, Markert U . Establishment of a one-sided ex vivo human placenta perfusion model to assess adhesion and invasion behavior of T cell leukemia cell lines. Leuk Lymphoma. 2012; 54(8):1811-3. DOI: 10.3109/10428194.2012.758844. View

17.

Smith J, Gaikwad A, Mosley S, Coffer 2nd L, Cegelski J, Alcorn J . Utilization of an ex vivo human placental perfusion model to predict potential fetal exposure to carboplatin during pregnancy. Am J Obstet Gynecol. 2013; 210(3):275.e1-9. DOI: 10.1016/j.ajog.2013.12.013. View

18.

Kusuma G, Manuelpillai U, Abumaree M, Pertile M, Brennecke S, Kalionis B . Mesenchymal stem cells reside in a vascular niche in the decidua basalis and are absent in remodelled spiral arterioles. Placenta. 2015; 36(3):312-21. DOI: 10.1016/j.placenta.2014.12.014. View

19.

Squillaro T, Peluso G, Galderisi U . Clinical Trials With Mesenchymal Stem Cells: An Update. Cell Transplant. 2015; 25(5):829-48. DOI: 10.3727/096368915X689622. View

20.

Xie J, Peng C, Zhao Q, Wang X, Yuan H, Yang L . Osteogenic differentiation and bone regeneration of iPSC-MSCs supported by a biomimetic nanofibrous scaffold. Acta Biomater. 2015; 29:365-379. DOI: 10.1016/j.actbio.2015.10.007. View