Skip to main content
SpringerLink
Log in

Simultaneous isolation of vascular endothelial cells and mesenchymal stem cells from the human umbilical cord

  • Report
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Abstract

The umbilical cord represents the link between mother and fetus during pregnancy. This cord is usually discarded as a biological waste after the child’s birth; however, its importance as a “store house” of stem cells has been explored recently. We developed a method of simultaneous isolation of endothelial cells (ECs) from the vein and mesenchymal stem cells from umbilical cord Wharton’s jelly of the same cord. The isolation protocol has been simplified, modified, and improvised with respect to choice of enzyme and enzyme mixture, digestion time, cell yield, cell growth, and culture medium. Isolated human umbilical vascular ECs (hUVECs) were positive for von-Willibrand factor, a classical endothelial marker, and could form capillary-like structures when seeded on Matrigel, thus proving their functionality. The isolated human umbilical cord mesenchymal stem cells (hUCMSCs) were found positive for CD44, CD90, CD 73, and CD117 and were found negative for CD33, CD34, CD45, and CD105 surface markers; they were also positive for cytoskeleton markers of smooth muscle actin and vimentin. The hUCMSCs showed multilineage differentiation potential and differentiated into adipogenic, chondrogenic, osteogenic, and neuronal lineages under influence of lineage specific differentiation medium. Thus, isolating endothelial cells as well as mesenchymal cells from the same umbilical cord could lead to complete utilization of the available tissue for the tissue engineering and cell therapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1.
Figure 2.
Figure 3.

References

  • Chao K. C.; Chao K. F.; Fu Y. S.; Liu S. H. Islet like clusters derived from mesenchymal stem cells in Wharton’s jelly of human umbilical cord for transplantation to control type 1 diabetes. PLoS One 1: e1451; 2008. doi:10.1371/journal.pone.0001451.

    Article  Google Scholar 

  • Dales J.; Garcia S.; Bonnier P.; Duffaud F.; Andrac-Meyer L.; Ramuz O.; Lavaut M.; Allasia C.; Charpin C. CD105 expression is a marker of high metastatic risk and poor outcome in breast carcinomas. Correlations between immunohistochemical analysis and long-term follow-up in a series of 929 patients. Am. J. Clin. Pathol. 1193: 374–380; 2003. doi:10.1309/1KF54L6RB625556W.

    Article  PubMed  Google Scholar 

  • Erices A.; Conget P.; Minguell J. Mesenchymal progenitor cells in human umbilical cord blood. Br. J. Haematol. 109: 235–242; 2000. doi:10.1046/j.1365-2141.2000.01986.x.

    Article  PubMed  CAS  Google Scholar 

  • Fonsatti E.; Sigalotti L.; Arslan P.; Altomonte M.; Maio M. Emerging role of endoglin (cd105) as a marker of angiogenesis with clinical potential in human malignancies. Current Cancer Drug Targets 36: 427–432; 2003. doi:10.2174/1568009033481741.

    Article  PubMed  CAS  Google Scholar 

  • Hayani A.; Lampeter E.; Viswanatha D.; Morgan D.; Salvi S. N. First report of autologous cord blood transplantation in the treatment of child with leukemia. Pediatrics 119: 296–300; 2007. doi:10.1542/peds.2006-1009.

    Article  Google Scholar 

  • Jaffe E. A.; Nachman R. L.; Becker C. G.; Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J. Clin. Invest. 5211: 1973; 2745–2756. doi:10.1172/JCI107470.

    Google Scholar 

  • Karahuseyinoglu S.; Cinar O.; Kilic E.; Kara F.; Akay G.; Demiralp D.; Tukun A.; Uckan D.; Can A. Biology of stem cells in human umbilical cord stroma: in situ and in vitro surveys. Stem. Cells 22: 617–624; 2004. doi:10.1634/stemcells.22-4-617.

    Article  Google Scholar 

  • Ko Y.; Totzke G.; Seewald S.; Schmitz U.; Schiermeyer B.; Meyer Z. U.; Brickwedde M. K.; Vetter H.; Sachinidis A. Native low density lipoprotein (LDL) includes the expression of the early growth response gene-1 in human umbilical arterial endothelial cells. Eur. J. Cell. Biol. 683: 306–302; 1995.

    PubMed  CAS  Google Scholar 

  • Laughlin M. J.; Barker J.; Bambach B.; Koc O. N.; Rizzieri D. A.; Wagner J. E.; Gerson S. L.; Lazarus H. M.; Cairo M.; Stevens C. E.; Rubinstein P.; Kurtzberg J. Hematopoietic engraftment and survival in adult recipient of umbilical cord blood from unrelated donors. N. Engl. J Med. 344: 1851–1822; 2001. doi:10.1056/NEJM200106143442402.

    Article  Google Scholar 

  • Lee O. K.; Kuo T. K.; Chen W. M.; Lee K. D.; Hsieh S. L.; Chen T. H. Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood 103: 1669–1675; 2004. doi:10.1182/blood-2003-05-1670.

    Article  PubMed  CAS  Google Scholar 

  • Lu-Lu L.; Yong Jun L.; Shao-Guang Y.; Quin-jun Z.; Xin W.; Wei G.; Zhi-Bo H.; Zhen-Shu X.; Yong-Xin L.; Delong L.; Zhi-Zhe C.; Zhong-Chao H. Isolation and characterization of human umbilical cord mesenchymal stem cells with hematopoiesis-supportive function and other potentials. Haematologica 91: 1017–1026; 2006.

    Google Scholar 

  • Mareschi K.; Biasin E.; Piacibello W.; Aglietta M.; Madon E.; Fagioli F. Isolation of human mesenchymal stem cells: bone marrow verses umbilical cord blood. Haematologica 86: 1099–1100; 2001.

    PubMed  CAS  Google Scholar 

  • Mitchell K. E.; Weiss M. L.; Mitchell B. M.; Martin P.; Davis D.; Morales L.; Helwig B.; Beerenstrauch M.; Abou-Easa K.; Hildreth T.; Troyer D. Matrix from Wharton’s jelly from neurons and glia. Stem Cells 21: 50–60; 2003.

    PubMed  CAS  Google Scholar 

  • Nanaev A. K.; Kohnen G.; Milovanov A. P.; Domogatsky S. P.; Kaufmann P. Stromal differentiation and architecture of the human umbilical cord. Placenta 18: 53–64; 1997. doi:10.1016/S0143-4004(97)90071-0.

    Article  PubMed  CAS  Google Scholar 

  • Phadnis S. M.; Joglekar M. V.; Venkateshan V.; Ghaskadbi S. M.; Hardikar A. A.; Bhonde R. R. Human umbilical cord blood serum promotes growth, proliferation as well as differentiation of human bone marrow-derived progenitor cells. In Vitro Cell Dev. Biol.—Animal 4210: 283–286; 2006.

    Google Scholar 

  • Romanov Y. A.; Svintsitskaya V. A.; Smirnov V. N. Searching for alternative sources of postnatal human mesenchymal stem cells candidate MSC-like cells from umbilical cord. Stem Cells 21: 105–110; 2003. doi:10.1634/stemcells.21-1-105.

    Article  PubMed  Google Scholar 

  • Secco M.; Zucconi E.; Vieira N. M.; Fogaca L. L. Q.; Cerqueira A.; Carvalho M. D. F.; Jazededje T.; Okamoto O. K.; Muotri A. R.; Zatz M. Multipotent stem cells from umbilical cord: cord is richer than blood!. Stem Cells 261: 146–50; 2007. doi:10.1634/stemcells.2007-0381.

    Article  PubMed  Google Scholar 

  • Troyer D. L.; Weiss M. L. Concise review: Wharton’s jelly-derived cells are a primitive stromal cell population. Stem Cells 26: 591–599; 2008. doi:10.1634/stemcells.2007-0439.

    Article  PubMed  Google Scholar 

  • Ulrich-Merzenich G.; Metzner C.; Bhonde R. R.; Malsch G.; Schiermeyer B.; Vetter H. Simultaneous isolation of Endothelial and smooth muscle cells from human umbilical artery or vein and their growth response to low density lipoproteins. In Vitro Cell Dev. Biol.—Animal 38: 265–272; 2002. doi:10.1290/1071-2690(2002)038<0265:SIOEAS>2.0.CO;2.

    Article  Google Scholar 

  • Wang H. S.; Hung S. C.; Peng S. T.; Huang C. C.; Wei H. M.; Guo Y. J.; Fu Y. S.; Lai M. C.; Chen C. C. Mesenchymal stem cells in the Wharton’s jelly of human umbilical cord. Stem Cells 22: 1330–1337; 2004. doi:10.1634/stemcells.2004-0013.

    Article  PubMed  Google Scholar 

  • William, P. L.; Bannister, L. H.; Berry, M. M.; Collins, P.; Dyson, M.; Dussek, J. E.; Fergusson, M. W. J. Gray’s anatomy, 38th edn. ELBS Churchill Livingstone, London; 1995.

Download references

Acknowledgments

Authors wish to thank Director, NCCS for providing necessary facilities. Thanks are also due to Dr. Meeta Nakhare, Head of the Gynecology Department, Ratna Memorial Hospital, Pune, India for human umbilical cord samples and Department of Biotechnology, Government of India for providing financial support to carry out this work.

Author information

Authors and Affiliations

  1. National Center for Cell Science, Ganeshkhind, Pune, 411007, India

    Sachin S. Kadam, Shubha Tiwari & Ramesh R. Bhonde

Authors
  1. Sachin S. Kadam
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shubha Tiwari
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ramesh R. Bhonde
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ramesh R. Bhonde.

Additional information

Editor: J. Denry Sato

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kadam, S.S., Tiwari, S. & Bhonde, R.R. Simultaneous isolation of vascular endothelial cells and mesenchymal stem cells from the human umbilical cord. In Vitro Cell.Dev.Biol.-Animal 45, 23–27 (2009). https://doi.org/10.1007/s11626-008-9155-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-008-9155-4

Keywords

Search

Navigation