Chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells in vitro

ABSTRACT Different therapeutic techniques have been developed for regeneration of articular cartilage injuries, but none has provided an optimal solution to their treatment. Human umbilical cord blood‐mesenchymal Stem Cells (HUCB‐MSCs) have been considered as promising alternative cell source for ca...

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Published in:	Microscopy research and technique Vol. 78; no. 8; pp. 667 - 675
Main Authors:	Mostafa Ibrahim, Azza, Mohamed Elgharabawi, Nesrine, Mohamed Makhlouf, Manal, Yahia Ibrahim, Omnia
Format:	Journal Article
Language:	English
Published:	United States Blackwell Publishing Ltd 01-08-2015 Wiley Subscription Services, Inc
Subjects:	Adult Cartilage Cell Differentiation - physiology Cells, Cultured Chondrocytes - chemistry Chondrocytes - cytology Chondrocytes - metabolism Chondrogenesis - physiology chondrogenic differentiation Differentiation Female Gene expression HUCB Human Human Umbilical Vein Endothelial Cells - chemistry Human Umbilical Vein Endothelial Cells - cytology Human Umbilical Vein Endothelial Cells - metabolism Humans Immunophenotyping In vitro testing Male Mesenchymal Stromal Cells - chemistry Mesenchymal Stromal Cells - cytology Mesenchymal Stromal Cells - metabolism Middle Aged MSCs Optimization Reverse Transcriptase Polymerase Chain Reaction RT-PCR Stem cells Umbilical cords Young Adult chondrogenic differentiation MSCs RT-PCR HUCB
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Summary:	ABSTRACT Different therapeutic techniques have been developed for regeneration of articular cartilage injuries, but none has provided an optimal solution to their treatment. Human umbilical cord blood‐mesenchymal Stem Cells (HUCB‐MSCs) have been considered as promising alternative cell source for cartilage repair. Objectives: Examining the success rate of MSCs isolation from HUCB as well as chondrogenic differentiation potential of HUCB‐MSCs in vitro. Materials and methods: 32 UCB samples were collected, in addition to 5 bone marrow (BM) and 5 peripheral blood (PB) samples, taken as reference controls. Samples were used for mononuclear cells isolation from which MSCs were expanded under complete aseptic conditions, were verified morphologically and through the presence of CD44 and CD105, and absence of CD34. Results: Success rate of UCB‐MSCs isolation was (25%), a rate that was lower than those of PB (40%) and BM (80%). Accordingly, certain input parameters have been recommended for successful MSCs isolation from UCB. On selecting samples in which recommended parameters were fulfilled, success rate was increased to 72%. This was together with providing optimal experiment conditions; mainly type of expansion medium, success rate reached 80%. Then, successfully expanded MSCs were subjected to chondrogenic differentiation by culturing in pelleted micromass system in presence of transforming growth factor beta‐1 and chondrogenic medium devoid of fetal bovine serum to evaluate their ability to undergo chondrogenesis. Differentiation was verified microscopically using special stains, and proved by reverse transcriptase‐polymerase chain reaction for expression of aggrecan and collagen II genes. In conclusion, in vitro differentiation into chondrocytes is possible from HUCB‐MSCs. Microsc. Res. Tech. 78:667–675, 2015. © 2015 Wiley Periodicals, Inc.
Bibliography:	Faculty of Medicine, El-Kasr El-Aini Hospital, Cairo University, Cairo, Egypt ArticleID:JEMT22520 istex:B95FCE940B3E9EEA171D9A870149D8E54A2088B3 ark:/67375/WNG-GF8X0M2P-8 REVIEW EDITOR: Prof. Alberto Diaspro ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1059-910X 1097-0029
DOI:	10.1002/jemt.22520