Effects of basic fibroblast growth factor in culture of mesenchymal stem cells derived from Wharton's jelly of human umbilical cord

Yun Tao Ba, Fangxia Guan, Hu Xiang, Yang Bo, Du Ying, Tian Xiang Zhang, Tian Yi, Xiao Jun Qiao, Cheng Chun Wang, Chen Xi Gu, Ning Jing Lei, Xiao Wei Wang

Research output: Contribution to journalArticle

Abstract

BACKGROUND: During culture of mesenchymal stem cells (MSCs) derived from Wharton's jelly of human umbilical cord. MSC morphology tends to become hypertrophic and irregular. MSCs were found to have higher rate of death and not be adapt to be adherent after passage. It is necessary to find a method to maintain its stability. OBJECTIVE: To isolate MSCs from human umbilical cord wharton's jelly by tissue block method, and to investigate basic fibroblast growth factor (bFGF) effects on biological characteristics of MSCs. METHODS: Human umbilical cord mesenchymal stem cells (hUCMSCs) were separated by tissue block method. Tissue fragments in control group were cultured in growth medium consisting of Dulbecco's Modified Essential /F12 Media (DMEM/F12) and 10% volume fraction of fetal bovine serum. Tissue fragments in experiment group were cultured in growth medium including 20 ug/L bFGF besides these in control group. Time of tissue blocks emigrating from cells and morphology of cells were observed. The medium was changed every 3-4 days. When the cells reached 80%-90% confluency, they were detached with 0.25% trypsin and were passaged at a ratio of 1: 2 or 1:3. RESULTS AND CONCLUSION: A few long spindle or flat fibroblast-like cells were presented firstly after 8-10 days of incubation in control group, while in experiment group it took 6-8 days. One week later, lots of long spindle or flat fibroblast-like cells like whirlpool around micro-mass were presented in two groups. In the first 3 passages, cell morphous were similar and cells were passaged at approximately equivalent time. After passage 3, cells in experiment group were easier to be adherent, lower rate of death and better proliferation ability (shown by growth curve) in comparison with these in control group. Flow cytometry revealed that cell cycle at the passages 3 and 6 showed the percentage of G0/G1 was more than 70% respectively. CD44 and CD29 were highly expressed on the surface of passages 3 and 6 cells, whereas the expression of HLA-ABC was less positive, but there was negative for CD34, CD45 and HLA-DR. Cells from Wharton's jelly of the human umbilical cords, which have the biological characteristics of MSCs, have been isolated by tissue culture method. bFGF can shorten the time that MSCs are presented firstly, improve its adherence and proliferation ability and maintain its morphological stability in some degree, moreover keep surface marker expression stability of MSCs.

Original languageEnglish (US)
Pages (from-to)3513-3517
Number of pages5
JournalJournal of Clinical Rehabilitative Tissue Engineering Research
Volume14
Issue number19
DOIs
StatePublished - May 7 2010
Externally publishedYes

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Wharton Jelly
Umbilical Cord
Fibroblast Growth Factor 2
Fibroblasts
Stem cells
Mesenchymal Stromal Cells
Cell culture
Tissue
Control Groups
Growth
Intercellular Signaling Peptides and Proteins
Tissue culture
Flow cytometry
Experiments
HLA-DR Antigens
Mortality
Trypsin
Volume fraction
Cell Cycle
Flow Cytometry

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biomedical Engineering
  • Transplantation

Cite this

Effects of basic fibroblast growth factor in culture of mesenchymal stem cells derived from Wharton's jelly of human umbilical cord. / Ba, Yun Tao; Guan, Fangxia; Xiang, Hu; Bo, Yang; Ying, Du; Zhang, Tian Xiang; Yi, Tian; Qiao, Xiao Jun; Wang, Cheng Chun; Gu, Chen Xi; Lei, Ning Jing; Wang, Xiao Wei.

In: Journal of Clinical Rehabilitative Tissue Engineering Research, Vol. 14, No. 19, 07.05.2010, p. 3513-3517.

Research output: Contribution to journalArticle

Ba, Yun Tao ; Guan, Fangxia ; Xiang, Hu ; Bo, Yang ; Ying, Du ; Zhang, Tian Xiang ; Yi, Tian ; Qiao, Xiao Jun ; Wang, Cheng Chun ; Gu, Chen Xi ; Lei, Ning Jing ; Wang, Xiao Wei. / Effects of basic fibroblast growth factor in culture of mesenchymal stem cells derived from Wharton's jelly of human umbilical cord. In: Journal of Clinical Rehabilitative Tissue Engineering Research. 2010 ; Vol. 14, No. 19. pp. 3513-3517.
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title = "Effects of basic fibroblast growth factor in culture of mesenchymal stem cells derived from Wharton's jelly of human umbilical cord",
abstract = "BACKGROUND: During culture of mesenchymal stem cells (MSCs) derived from Wharton's jelly of human umbilical cord. MSC morphology tends to become hypertrophic and irregular. MSCs were found to have higher rate of death and not be adapt to be adherent after passage. It is necessary to find a method to maintain its stability. OBJECTIVE: To isolate MSCs from human umbilical cord wharton's jelly by tissue block method, and to investigate basic fibroblast growth factor (bFGF) effects on biological characteristics of MSCs. METHODS: Human umbilical cord mesenchymal stem cells (hUCMSCs) were separated by tissue block method. Tissue fragments in control group were cultured in growth medium consisting of Dulbecco's Modified Essential /F12 Media (DMEM/F12) and 10{\%} volume fraction of fetal bovine serum. Tissue fragments in experiment group were cultured in growth medium including 20 ug/L bFGF besides these in control group. Time of tissue blocks emigrating from cells and morphology of cells were observed. The medium was changed every 3-4 days. When the cells reached 80{\%}-90{\%} confluency, they were detached with 0.25{\%} trypsin and were passaged at a ratio of 1: 2 or 1:3. RESULTS AND CONCLUSION: A few long spindle or flat fibroblast-like cells were presented firstly after 8-10 days of incubation in control group, while in experiment group it took 6-8 days. One week later, lots of long spindle or flat fibroblast-like cells like whirlpool around micro-mass were presented in two groups. In the first 3 passages, cell morphous were similar and cells were passaged at approximately equivalent time. After passage 3, cells in experiment group were easier to be adherent, lower rate of death and better proliferation ability (shown by growth curve) in comparison with these in control group. Flow cytometry revealed that cell cycle at the passages 3 and 6 showed the percentage of G0/G1 was more than 70{\%} respectively. CD44 and CD29 were highly expressed on the surface of passages 3 and 6 cells, whereas the expression of HLA-ABC was less positive, but there was negative for CD34, CD45 and HLA-DR. Cells from Wharton's jelly of the human umbilical cords, which have the biological characteristics of MSCs, have been isolated by tissue culture method. bFGF can shorten the time that MSCs are presented firstly, improve its adherence and proliferation ability and maintain its morphological stability in some degree, moreover keep surface marker expression stability of MSCs.",
author = "Ba, {Yun Tao} and Fangxia Guan and Hu Xiang and Yang Bo and Du Ying and Zhang, {Tian Xiang} and Tian Yi and Qiao, {Xiao Jun} and Wang, {Cheng Chun} and Gu, {Chen Xi} and Lei, {Ning Jing} and Wang, {Xiao Wei}",
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T1 - Effects of basic fibroblast growth factor in culture of mesenchymal stem cells derived from Wharton's jelly of human umbilical cord

AU - Ba, Yun Tao

AU - Guan, Fangxia

AU - Xiang, Hu

AU - Bo, Yang

AU - Ying, Du

AU - Zhang, Tian Xiang

AU - Yi, Tian

AU - Qiao, Xiao Jun

AU - Wang, Cheng Chun

AU - Gu, Chen Xi

AU - Lei, Ning Jing

AU - Wang, Xiao Wei

PY - 2010/5/7

Y1 - 2010/5/7

N2 - BACKGROUND: During culture of mesenchymal stem cells (MSCs) derived from Wharton's jelly of human umbilical cord. MSC morphology tends to become hypertrophic and irregular. MSCs were found to have higher rate of death and not be adapt to be adherent after passage. It is necessary to find a method to maintain its stability. OBJECTIVE: To isolate MSCs from human umbilical cord wharton's jelly by tissue block method, and to investigate basic fibroblast growth factor (bFGF) effects on biological characteristics of MSCs. METHODS: Human umbilical cord mesenchymal stem cells (hUCMSCs) were separated by tissue block method. Tissue fragments in control group were cultured in growth medium consisting of Dulbecco's Modified Essential /F12 Media (DMEM/F12) and 10% volume fraction of fetal bovine serum. Tissue fragments in experiment group were cultured in growth medium including 20 ug/L bFGF besides these in control group. Time of tissue blocks emigrating from cells and morphology of cells were observed. The medium was changed every 3-4 days. When the cells reached 80%-90% confluency, they were detached with 0.25% trypsin and were passaged at a ratio of 1: 2 or 1:3. RESULTS AND CONCLUSION: A few long spindle or flat fibroblast-like cells were presented firstly after 8-10 days of incubation in control group, while in experiment group it took 6-8 days. One week later, lots of long spindle or flat fibroblast-like cells like whirlpool around micro-mass were presented in two groups. In the first 3 passages, cell morphous were similar and cells were passaged at approximately equivalent time. After passage 3, cells in experiment group were easier to be adherent, lower rate of death and better proliferation ability (shown by growth curve) in comparison with these in control group. Flow cytometry revealed that cell cycle at the passages 3 and 6 showed the percentage of G0/G1 was more than 70% respectively. CD44 and CD29 were highly expressed on the surface of passages 3 and 6 cells, whereas the expression of HLA-ABC was less positive, but there was negative for CD34, CD45 and HLA-DR. Cells from Wharton's jelly of the human umbilical cords, which have the biological characteristics of MSCs, have been isolated by tissue culture method. bFGF can shorten the time that MSCs are presented firstly, improve its adherence and proliferation ability and maintain its morphological stability in some degree, moreover keep surface marker expression stability of MSCs.

AB - BACKGROUND: During culture of mesenchymal stem cells (MSCs) derived from Wharton's jelly of human umbilical cord. MSC morphology tends to become hypertrophic and irregular. MSCs were found to have higher rate of death and not be adapt to be adherent after passage. It is necessary to find a method to maintain its stability. OBJECTIVE: To isolate MSCs from human umbilical cord wharton's jelly by tissue block method, and to investigate basic fibroblast growth factor (bFGF) effects on biological characteristics of MSCs. METHODS: Human umbilical cord mesenchymal stem cells (hUCMSCs) were separated by tissue block method. Tissue fragments in control group were cultured in growth medium consisting of Dulbecco's Modified Essential /F12 Media (DMEM/F12) and 10% volume fraction of fetal bovine serum. Tissue fragments in experiment group were cultured in growth medium including 20 ug/L bFGF besides these in control group. Time of tissue blocks emigrating from cells and morphology of cells were observed. The medium was changed every 3-4 days. When the cells reached 80%-90% confluency, they were detached with 0.25% trypsin and were passaged at a ratio of 1: 2 or 1:3. RESULTS AND CONCLUSION: A few long spindle or flat fibroblast-like cells were presented firstly after 8-10 days of incubation in control group, while in experiment group it took 6-8 days. One week later, lots of long spindle or flat fibroblast-like cells like whirlpool around micro-mass were presented in two groups. In the first 3 passages, cell morphous were similar and cells were passaged at approximately equivalent time. After passage 3, cells in experiment group were easier to be adherent, lower rate of death and better proliferation ability (shown by growth curve) in comparison with these in control group. Flow cytometry revealed that cell cycle at the passages 3 and 6 showed the percentage of G0/G1 was more than 70% respectively. CD44 and CD29 were highly expressed on the surface of passages 3 and 6 cells, whereas the expression of HLA-ABC was less positive, but there was negative for CD34, CD45 and HLA-DR. Cells from Wharton's jelly of the human umbilical cords, which have the biological characteristics of MSCs, have been isolated by tissue culture method. bFGF can shorten the time that MSCs are presented firstly, improve its adherence and proliferation ability and maintain its morphological stability in some degree, moreover keep surface marker expression stability of MSCs.

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