Simultaneous isolation of vascular endothelial cells and mesenchymal stem cells from the human umbilical cord
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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.
KeywordsUmbilical cord vein Human umbilical cord vascular endothelial cells Wharton’s jelly Human umbilical cord matrix stem cells
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.
- 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.PubMedCrossRefGoogle 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.PubMedGoogle 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.CrossRefGoogle 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
- 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
- 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.PubMedCrossRefGoogle 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.CrossRefGoogle 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.Google Scholar