IN VITRO EVALUATION OF CHONDROCYTES COMBINED WITH Wharton's JELLY OF HUMAN UMBILICAL CORD ORIENTED SCAFFOLD_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

LüHanning ¹ , XUGuojiao ¹ , GAIYuxin ¹ , CHENLi ¹ , LIUShuyun ² , ZHAOPeng ¹ , LUShibi ² , ZHANGLi ² ,  GUOQuanyi ² ,  YANGJianhua ¹

1. Department of Orthopedics, the First Affiliated Hospital of Jiamusi University, Jiamusi Heilongjiang, 154002, P. R. China;
2. Institute of Orthopedics, Chinese PLA General Hospital;

Corresponding?author：

GUOQuanyi, Email: doctorguo@163.com; YANGJianhua, Email: jianhua01@163.com

Keywords：

Cartilage tissue engineering; Wharton's jelly of human umbilical cord; Oriented scaffold; Chondrocytes

DOI：

10.7507/1002-1892.20140223

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective To assess the role and effect of Wharton's jelly of human umbilical cord oriented scaffold on chondrocytes co-cultured in vitro. Methods Chondrocytes from shoulder cartilage of adult New Zealand rabbits were isolated,cultured,amplified,and labelled using fluorescent dye PKH26.Cells were extracted from human umbilical cord tissue using wet-grinding chemical technology to prepare the Wharton's jelly of human umbilical cord oriented scaffold by freeze-drying and cross-linking technology.Second generation of chondrocytes were cultured with Wharton's jelly of human umbilical cord oriented scaffold.Inverted microscope and scanning electron microscope (SEM) were used to observe the cell distribution and adhesion on the scaffold; extracellular matrix secretion of the chondrocytes were observed by toluidine blue and safranin O staining.Cells distribution and proliferation on the scaffold were assessed by fluorescein diacetate-propidium iodide (FDA-PI) and Hoechst33258 staining.The viability of the in vitro cultured and PKH26 fluorescence labelled chondrocytes on the scaffold were assessed via fluorescence microscope. Results Inverted microscope showed that the cells cultured on the scaffold for 3 days were round or oval shaped and evenly distributed into space of the scaffold.SEM observation showed that large number of cultured cells adhered to the pores between the scaffolds and were round or oval shape,which aggregated,proliferated,and arranged vertically on longitudinally oriented scaffold at 7 days after culture.Histological observation showed that cells distributed and proliferated on the scaffold,and secreted large amount of extracellular matrix at 7 days.Scaffold could guide cell migration and proliferation,and could effectively preserve and promote the secretion of extracellular matrix.Cell viability assessments at 3 days after culture showed most of the adhered cells were living and the viability was more than 90%.PKH26 labelled chondrocytes were seen,which distributed uniformly along the pore of oriented scaffold,and exuberantly proliferated. Conclusion Wharton's jelly of human umbilical cord oriented scaffold favors adhesion,proliferation,and survival of chondrocytes.It possesses a favorable affinity and cell compatibility.Thus,it is an ideal scaffold for cartilage tissue engineering.

Citation： LüHanning, XUGuojiao, GAIYuxin, CHENLi, LIUShuyun, ZHAOPeng, LUShibi, ZHANGLi, GUOQuanyi, YANGJianhua. IN VITRO EVALUATION OF CHONDROCYTES COMBINED WITH Wharton's JELLY OF HUMAN UMBILICAL CORD ORIENTED SCAFFOLD. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(8): 1017-1022. doi: 10.7507/1002-1892.20140223 Copy

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10.	Liu S,Hou KD,Yuan M,et al.Characteristics of mesenchymal stem cells derived from Wharton's Jelly of human umbilical cord and for fabrication of non-scaffold tissue-engineered cartilage.J Biosci Bioeng,2014,117(2):229-235.
11.	Detamore MS.Human umbilical cord mesenchymal stromal cells in regenerative medicine.Stem Cell Res Ther,2013,4(6):142.
12.	Liu S,Yuan M,Hou K,et al.Immune characterization of mesenchymal stem cells in human umbilical cord Wharton's Jelly and derived cartilage cells.Cell Immunol,2012,278(1-2):35-44.
13.	趙鵬,劉舒云,盧世璧,等.軟骨細胞外基質對臍帶Wharton膠間充質干細胞生物學性狀的作用.中國矯形外科雜志,2013,21(11):1127-1132.

1. Baba K,Yamazaki Y,Ishiguro M,et al.Osteogenic potential of human umbilical cord-derived mesenchymal stromal cells cultured with umbilical cord blood-derived fibrin:a preliminary study.J Craniomaxillofac Surg,2013,41(8):775-782.
2. Feng G,Wan Y,Balian G,et al.Adenovirus-mediated expression of growth and differentiation factor-5 promotes chondrogenesis of adipose stem cells.Growth Factors,2008,26(3):132-142.
3. Corrao S,La Rocca G,Lo Iacono M,et al.Umbilical cord revisited:from Wharton's jelly myofibroblasts to mesenchymal stem cells.Histol Histopathol,2013,28(10):1235-1244.
4. Fong CY,Gauthaman K,Cheyyatraivendran S,et al.Human umbilical cord Wharton's jelly stem cells and its conditioned medium support hematopoietic stem cell expansion ex vivo.J Cell Biochem,2012,113(2):658-668.
5. 許海委,徐寶山,楊強,等.細胞共培養法誘導兔脂肪來源干細胞向軟骨細胞分化的實驗研究.中國修復重建外科雜志,2013,27(2):193-198.
6. Nagamura-Inoue T,He H.Umbilical cord-derived mesenchymal stem cells:Their advantages and potential clinical utility.World J Stem Cells,2014,6(2):195-202.
7. 柳箐,宇麗,許超,等.長期培養人臍帶間充質干細胞分化能力改變的研究.中國生物工程雜志,2012,32(2):8-16.
8. Wang L,Zhao L,Detamore MS.Human umbilical cord mesenchymal stromal cells in a sandwich approach for osteochondral tissue engineering.J Tissue Eng Regen Med,2011,5(9):712-721.
9. Romanowicz L,Bańkowski E.Lipid compounds of human Wharton's jelly and their alterations in preeclampsia.Int J Exp Pathol,2010,91(1):1-9.
10. Liu S,Hou KD,Yuan M,et al.Characteristics of mesenchymal stem cells derived from Wharton's Jelly of human umbilical cord and for fabrication of non-scaffold tissue-engineered cartilage.J Biosci Bioeng,2014,117(2):229-235.
11. Detamore MS.Human umbilical cord mesenchymal stromal cells in regenerative medicine.Stem Cell Res Ther,2013,4(6):142.
12. Liu S,Yuan M,Hou K,et al.Immune characterization of mesenchymal stem cells in human umbilical cord Wharton's Jelly and derived cartilage cells.Cell Immunol,2012,278(1-2):35-44.
13. 趙鵬,劉舒云,盧世璧,等.軟骨細胞外基質對臍帶Wharton膠間充質干細胞生物學性狀的作用.中國矯形外科雜志,2013,21(11):1127-1132.

Chinese Journal of Reparative and Reconstructive Surgery

IN VITRO EVALUATION OF CHONDROCYTES COMBINED WITH Wharton's JELLY OF HUMAN UMBILICAL CORD ORIENTED SCAFFOLD

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