DIFFERENTIATING INTO ISLET-LIKE CELL CLUSTERS FROM HUMAN FIRST TRIMESTER UMBILICAL CORD PERIVASCULAR CELLS IN VITRO_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

 CAOMei ¹ , ZHANGJianbo ¹ ,  YEShangmian ¹ , LIHao ² , ZHAOJian ²

1. Core Laboratory, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu Sichuan, 610072, P. R. China;
2. College of Life Sciences, Sichuan University;

Corresponding author：

CAOMei, Email: cm1023@163.com; YEShangmian, Email: shangmian.yie@gmail.com

Keywords：

First trimester; Human umbilical cord perivascular cells; Embryoid body; Islet-like cell; Insulin

DOI：

10.7507/1002-1892.20140250

Video：

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Objective To study the characteristics of the human umbilical cord perivascular cells (HUCPVC) isolated from human first trimester umbilical cord perivascular layer tissues and the differentiation into islet-like cell clusters in vitro. Methods The HUCPVC derived from human first trimester umbilical cord which was donated by the volunteers were isolated and subcultured. The surface markers such as stage-specific embryonic antigen 1 (SSEA-1), SSEA-3, SSEA-4, OCT-4, TRA-1-60, and TRA-1-81 were detected by immunohistochemical method. The first trimester HUCPVC were induced to embryoid bodies (EB)-like cell aggregations and islet-like cell clusters in vitro through a simple stepwise culture protocol (5 steps). The expressions of specific markers[α-fetoprotein (AFP), Nestin, and smooth muscle actin (SMA)] were measured by immunohistochemical method; and the ability of glucose-stimulated insulin secretion was analyzed. Results The first trimester HUCPVC were successfully isolated and could be passaged steadily more than 10 generations, which expressed SSEA-3, SSEA-4, OCT-4, TRA-1-61, and TRA-1-81. The first trimester HUCPVC were successfully induced into EB-like cell aggregations and islet-like cell clusters. The EB-like cell aggregations could express markers of three germ lineages:AFP, Nestin, and SMA. The islet-like cell clusters could release insulin significantly in response to elevated concentrations of glucose in vitro (t=7.444, P=0.002). The insulin contents were (23.2±5.3) mU/L and (7.0±0.5) mU/L in high and low glucose media, respectively. Conclusion The first trimester HUCPVC has the ability to differentiate into islet-like cell clusters which can secret insulin in vitro.

Citation： CAOMei, ZHANGJianbo, YEShangmian, LIHao, ZHAOJian. DIFFERENTIATING INTO ISLET-LIKE CELL CLUSTERS FROM HUMAN FIRST TRIMESTER UMBILICAL CORD PERIVASCULAR CELLS IN VITRO. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(9): 1156-1160. doi: 10.7507/1002-1892.20140250 Copy

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5.	Lumelsky N, Blondel O, Laeng P, et al.Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets.Science, 2001, 292(5520):1389-1394.
6.	Jiang W, Shi Y, Zhao D, et al.In vitro derivation of functional insulin-producing cells from human embryonic stem cells.Cell Res, 2007, 17(4):333-344.
7.	Lee SH, Song KS, Kim JY, et al.Novel thiophenyl C-aryl glucoside SGLT2 inhibitors as potential antidiabetic agents.Bioorg Med Chem, 2011, 19(19):5813-5832.
8.	Otake K, Azukizawa S, Takahashi K, et al.2-Acyl-tetrahydroisoquinoline-3-carboxylic acids:lead compounds with triple actions, peroxisome proliferator-activated receptor α/γ agonist and protein-tyrosine phosphatase 1B inhibitory activities.Chem Pharm Bull (Tokyo), 2011, 59(7):876-879.
9.	Shapiro AM, Lakey JR, Ryan EA, et al.Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen.N Engl J Med, 2000, 343(4):230-238.
10.	Matsumoto S, Tanaka K.Pancreatic islet cell transplantation using non-heart-beating donors (NHBDs).J Hepatobiliary Pancreat Surg, 2005, 12(3):227-230.
11.	Suva D, Garavaglia G, Menetrey J, et al.Non-hematopoietic human bone marrow contains long-lasting, pluripotential mesenchymal stem cells.J Cell Physiol, 2004, 198(1):110-118.
12.	Yu M, Xiao Z, Shen L, et al.Mid-trimester fetal blood-derived adherent cells share characteristics similar to mesenchymal stem cells but full-term umbilical cord blood does not.Br J Haematol, 2004, 124(5):666-675.
13.	Tsai MS, Lee JL, Chang YJ, et al.Isolation of human multipotent mesenchymal stem cells from second-trimester amniotic fluid using a novel two-stage culture protocol.Hum Reprod, 2004, 19(6):1450-1456.
14.	Guillot PV, Gotherstrom C, Chan J, et al.Human first-trimester fetal MSC express pluripotency markers and grow faster and have longer telomeres than adult MSC.Stem Cells, 2007, 25(3):646-654.
15.	Sarugaser R, Lickorish D, Baksh D, et al.Human umbilical cord perivascular (HUCPV) cells:a source of mesenchymal progenitors.Stem Cells, 2005, 23(2):220-229.
16.	Can A, Karahuseyinoglu S.Concise review:human umbilical cord stroma with regard to the source of fetus-derived stem cells.Stem Cells, 2007, 25(11):2886-2895.

1. Bongso A, Fong CY.The therapeutic potential, challenges and future clinical directions of stem cells from the Wharton's jelly of the human umbilical cord.Stem Cell Rev, 2013, 9(2):226-240.
2. Zebardast N, Lickorish D, Davies JE.Human umbilical cord perivascular cells (HUCPVC):A mesenchymal cell source for dermal wound healing.Organogenesis, 2010, 6(4):197-203.
3. Hong SH, Maghen L, Kenigsbcerg S, et al.Ontogeny of human umbilical cord perivascular cells:molecular and fate potential changes during gestation.Stem Cells Dev, 2013, 22(17):2425-2439.
4. Librach CL, Yie SM, Xiao R.Smmethod of isolation and use of cells derived from first trimester umbilical cord tissue.US, 20090074731[P].2009-03-19.
5. Lumelsky N, Blondel O, Laeng P, et al.Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets.Science, 2001, 292(5520):1389-1394.
6. Jiang W, Shi Y, Zhao D, et al.In vitro derivation of functional insulin-producing cells from human embryonic stem cells.Cell Res, 2007, 17(4):333-344.
7. Lee SH, Song KS, Kim JY, et al.Novel thiophenyl C-aryl glucoside SGLT2 inhibitors as potential antidiabetic agents.Bioorg Med Chem, 2011, 19(19):5813-5832.
8. Otake K, Azukizawa S, Takahashi K, et al.2-Acyl-tetrahydroisoquinoline-3-carboxylic acids:lead compounds with triple actions, peroxisome proliferator-activated receptor α/γ agonist and protein-tyrosine phosphatase 1B inhibitory activities.Chem Pharm Bull (Tokyo), 2011, 59(7):876-879.
9. Shapiro AM, Lakey JR, Ryan EA, et al.Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen.N Engl J Med, 2000, 343(4):230-238.
10. Matsumoto S, Tanaka K.Pancreatic islet cell transplantation using non-heart-beating donors (NHBDs).J Hepatobiliary Pancreat Surg, 2005, 12(3):227-230.
11. Suva D, Garavaglia G, Menetrey J, et al.Non-hematopoietic human bone marrow contains long-lasting, pluripotential mesenchymal stem cells.J Cell Physiol, 2004, 198(1):110-118.
12. Yu M, Xiao Z, Shen L, et al.Mid-trimester fetal blood-derived adherent cells share characteristics similar to mesenchymal stem cells but full-term umbilical cord blood does not.Br J Haematol, 2004, 124(5):666-675.
13. Tsai MS, Lee JL, Chang YJ, et al.Isolation of human multipotent mesenchymal stem cells from second-trimester amniotic fluid using a novel two-stage culture protocol.Hum Reprod, 2004, 19(6):1450-1456.
14. Guillot PV, Gotherstrom C, Chan J, et al.Human first-trimester fetal MSC express pluripotency markers and grow faster and have longer telomeres than adult MSC.Stem Cells, 2007, 25(3):646-654.
15. Sarugaser R, Lickorish D, Baksh D, et al.Human umbilical cord perivascular (HUCPV) cells:a source of mesenchymal progenitors.Stem Cells, 2005, 23(2):220-229.
16. Can A, Karahuseyinoglu S.Concise review:human umbilical cord stroma with regard to the source of fetus-derived stem cells.Stem Cells, 2007, 25(11):2886-2895.

Chinese Journal of Reparative and Reconstructive Surgery

DIFFERENTIATING INTO ISLET-LIKE CELL CLUSTERS FROM HUMAN FIRST TRIMESTER UMBILICAL CORD PERIVASCULAR CELLS IN VITRO

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