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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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

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.	许海委,徐宝山,杨强,等.细胞共培养法诱导兔脂肪来源干细胞向软骨细胞分化的实验研究.中国修复重建外科杂志,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,宇丽,许超,等.长期培养人脐带间充质干细胞分化能力改变的研究.中国生物工程杂志,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.	赵鹏,刘舒云,卢世璧,等.软骨细胞外基质对脐带Wharton胶间充质干细胞生物学性状的作用.中国矫形外科杂志,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. 许海委,徐宝山,杨强,等.细胞共培养法诱导兔脂肪来源干细胞向软骨细胞分化的实验研究.中国修复重建外科杂志,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,宇丽,许超,等.长期培养人脐带间充质干细胞分化能力改变的研究.中国生物工程杂志,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. 赵鹏,刘舒云,卢世璧,等.软骨细胞外基质对脐带Wharton胶间充质干细胞生物学性状的作用.中国矫形外科杂志,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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