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Histochemistry and Cell Biology

, Volume 138, Issue 4, pp 593–603 | Cite as

A dose-dependent function of follicular fluid on the proliferation and differentiation of umbilical cord mesenchymal stem cells (MSCs) of goat

  • Pubin Qiu
  • Yaofu Bai
  • Chao Liu
  • Xin He
  • Hui Cao
  • Mingzhao Li
  • Haijing Zhu
  • Jinlian HuaEmail author
Original Paper

Abstract

Umbilical cord (UC) has been suggested as a new source of mesenchymal stem cells (MSCs). In this report, we isolated MSCs from the fetal UC of goat and investigated their multipotency of differentiation into germ cells in vitro, in the presence of 0–20 % bovine follicular fluid (FF). The phenotypes, capacity of proliferation and expression of MSC markers were served as the indexes of multipotency of the isolated UC-MSCs, those were ascertained by growth curves, RT-PCR and immunofluorescent staining, respectively. Our results showed that the UC-MSCs shared a similar immunophenotype to those cells reported in mouse and human bone marrow MSCs, as well as some characteristics seen in embryonic stem cells (ESCs). In addition, our data also demonstrated that a dose-dependent function of FF on the states of differentiation of goat UC-MSCs. From 2 to 20 % of the FF can promote the proliferation of goat UC-MSC, especially the 5 % concentration of follicular fluid promote proliferation was significantly higher than 2 %. In contrast, higher concentration of follicular fluid (>10 %) induced goat UC-MSCs differentiation into oocyte-like cells. These findings provide an efficient model to study the mechanism on cell proliferation and germ cell differentiation in livestock using FF.

Keywords

Multipotent Mesenchymal stem cells (MSCs) Umbilical cord (UC) Germ cells Goat 

Notes

Acknowledgments

The authors sincerely thank for Dr. Xiaoming Liu to revise our manuscript. We appreciate the editor and reviewer’s excellent work and suggestions. This work was supported by grants from the Program (30972097) from National Natural Science Foundation of China, Key Program of State Education Ministry (109148), Program for New Century Excellent Talents in University (NCET-09-0654), Program of Shannxi Province (2011K02-06), The Fundamental Research Funds for the Central Universities (QN2011012), China Postdoctoral Science Foundation funded project (200801438).

Supplementary material

418_2012_975_MOESM1_ESM.tif (5.2 mb)
Supplemented Figure.1 The 3 month fetal goat ovary was used as positive control for immunofluorescence staining. The ovarian germ cells were positive for Dazl, Vasa, ZP2, ZP3, and Stra8, respectively, bar = 25 μm. Supplementary material 1 (TIFF 5302 kb)
418_2012_975_MOESM2_ESM.tif (3.1 mb)
Supplemented Figure.2 The cultured goat oocyte was positive for Vasa, Dazl, ZP2, ZP3, Stra8 and Scp3, bar = 200 μm. Supplementary material 2 (TIFF 3213 kb)
418_2012_975_MOESM3_ESM.tif (3.2 mb)
Supplemented Figure.3 The cultured goat UC-MSCs were negative for CD14, CD79a, HLA-DR; positive for CD105, CD73, and CD90, respectively. Supplementary material 3 (TIFF 3254 kb)
418_2012_975_MOESM4_ESM.tif (5.7 mb)
Supplemented Figure.4 Goat UC-MSCs differentiated into cells positive for oil red-O staining (adipocyte-like cells, A, Induced group, B, Control, bar = 200 μm) and alizarin red staining (osteoblast-like cells, C, Induced group, D, Control, bar = 200 μm), the expression of PPAR-γ (E) and OPN (Osteocalcin, F) were increased in induced goat UC-MSCs. G, UC-MSCs were induced differentiated into cells positive for alcian blue staining, bar = 100 μm. Supplementary material 4 (TIFF 5883 kb)
418_2012_975_MOESM5_ESM.tif (4.6 mb)
Supplemented Figure.5 The frequency of large round cells was increased with the increasing concentration of FF after culture in vitro, A, 2% FF (A). 5% FF (B), 10% FF (C), 20% FF (D), Control (0% FF, E). A-D, bar = 400 μm; E, bar = 200 μm. Supplementary material 5 (TIFF 4729 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Pubin Qiu
    • 1
  • Yaofu Bai
    • 1
  • Chao Liu
    • 1
  • Xin He
    • 1
  • Hui Cao
    • 1
    • 2
  • Mingzhao Li
    • 1
  • Haijing Zhu
    • 1
  • Jinlian Hua
    • 1
    Email author
  1. 1.College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering and Technology, Key Lab for Animal Biotechnology of Ministry of Agriculture of ChinaNorthwest A&F UniversityYanglingPeople’s Republic of China
  2. 2.Hope Bio-technology (Su Zhou) Co., LtdSu ZhouPeople’s Republic of China

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