Skip to main content

Advertisement

SpringerLink
Log in

In vitro trans-differentiation of human umbilical cord derived hematopoietic stem cells into hepatocyte like cells using combination of growth factors for cell based therapy

  • Original Research
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

The aim of the study was to develop a new strategy for the differentiation of hematopoietic stem cell (HSC) derived from UCB into hepatocyte like cells and also to estimate the effects of combination of fibroblast growth factor 4 (FGF 4) and hepatocyte growth factor (HGF) on hematopoietic stem cell differentiation. HSCs were isolated and purified by magnetic activated cell sorting. HSCs were induced to hepatocyte like cells under a 2-step protocol with combination of growth factors. Reverse transcription polymerase chain reaction was performed to detect multiple genes related to hepatocyte like cells development and function. Hepatocyte like morphology was illustrated by inverted repeat microscope and the secretion of albumin and α- fetoprotein by these cells was confirmed by enzyme linked immunosorbent assay. Hepatocyte like cells was observed at the end of the protocol (days 14). These differentiated cells were observed to show high expression of genes related to hepatocytes (tryptophan 2, 3-dioxygenase [TO], glucose 6-phosphate [G6P], cytokeratin 18 [CK 18], albumin and α- fetoprotein [AFP]). The quantities of albumin and AFP at day 0 were low and upon differentiation the cells were able to produce albumin and AFP at high levels. Our results show a new strategy for differentiation in a short duration, using a combination of growth factors for the differentiation of umbilical cord blood derived HSC into hepatocyte like cells under certain in vitro conditions. After further studies this approach has the potency, for widespread cell replacement therapy for liver diseases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Bromeyer HE (1995) Questions to be answered regarding umbilical cord blood hematopoietic stem and progenitor cells and their use in transplantation. Transfusion 35:694–702

    Article  Google Scholar 

  • Broxmeyer HE, Douglas GW, Hangoc G et al (1989) Human umbilical cord blood as a potential source of transplantable hematopoietic stem/progenitor cells. Proc Natl Acad Sci USA 86:3828–3832

    Article  CAS  Google Scholar 

  • Chomaczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:196–199

    Google Scholar 

  • Danet GH, Luongo JL, Butler G et al (2002) C1qRp defines a new human stem cell population with hematopoietic and hepatic potential. Proc Natl Acad Sci USA 99:10441–10445

    Article  CAS  Google Scholar 

  • Di Campli C, Piscaglia AC, Pierelli L et al (2004) A human umbilical cord stem cell rescue therapy in a murine model of toxic liver injury. Dig Liver Dis 36:603–613

    Article  CAS  Google Scholar 

  • Feng G, De-Quan W, Yah-Hua H et al (2008) Extracellular matrix gel is necessary for in vitro cultivation of insulin producing cells from human umbilical cord blood derived mesenchymal stem cells. Chinese Med J 121:811–818

    Google Scholar 

  • Fiegel HC, Lioznov MV (2003) Cortes-Dericks liver-specific gene expression in cultured human hematopoietic stem cells. Stem Cells 21:98–104

    Article  CAS  Google Scholar 

  • Forbes S, Vig P, Poulsom R et al (2002) Hepatic stem cells. J Pathol 197:510–518

    Article  Google Scholar 

  • Fox IJ, Chowdhury JR (2004) Hepatocyte transplantation. Amer J Transplant 4:7–13

    Article  CAS  Google Scholar 

  • Glimm H, Tang P, Clark-Lewis I et al (2002) Ex vivo treatment of proliferating human cord blood stem cells with stroma-derived factor-1 enhances their ability to engraft NOD/SCID mice. Blood 99:3454–3457

    Article  CAS  Google Scholar 

  • Gluckman E, Broxmeyer HE, Auerbach AD et al (1989) Hematopoietic reconstitution in a patient with Fanconi’s anemia by means of umbilical-cord blood from an HLA-identical sibling. N Engl J Med 321:1174–1178

    Article  CAS  Google Scholar 

  • Ishikawa F, Drake CJ, Yang S et al (2003) Transplanted human cord blood cells give rise to hepatocytes in engrafted mice. Ann NY Sci 996:174–185

    Article  CAS  Google Scholar 

  • Kurtzberg J, Laughlin M, Graham ML et al (1996) Placental blood as a source of hematopoietic stem cells for transplantation into unrelated recipients. N Engl J Med 335:157–166

    Article  CAS  Google Scholar 

  • Lazaro CA, Croager EJ, Mitchell C et al (2003) Establishment, characterization, and long-term maintenance of cultures of human fetal hepatocytes. Hepatology 38:1095–1106

    Article  Google Scholar 

  • Lee KD, Kuo TK, Whang-Peng J et al (2004) In vitro hepatic differentiation of human mesenchymal stem cells. Hepatology 40:1275–1284

    Article  CAS  Google Scholar 

  • Mayani H, Lansdorp PM (1998) Biology of human umbilical cord blood-derived hematopoietic stem/progenitor cells. Stem Cells 16:153–165

    Article  CAS  Google Scholar 

  • McAdams TA, Miller WM, Papoutsakis ET (1996) Hematopoietic cell culture therapies. Trends Biotechnol 14:388–396

    Article  CAS  Google Scholar 

  • Mossman T (1983) Rapid colorimetric assay of cellular growth and survival: a application to proliferation and cytotoxicity assays. J Immunol Method 65:55–63

    Article  Google Scholar 

  • Nakahata T, Ogawa M (1982) Hemopoietic colony-forming cells in umbilical cord blood with extensive capability to generate mono- and multipotential hemopoietic progenitors. J Clin Invest 70:1324–1328

    Article  CAS  Google Scholar 

  • Oh SH, Miyazaki M, Kouchi H et al (2000) Hepatocyte growth factor induces differentiation of adult rat bone marrow cells into a hepatocyte lineage in vitro. Biochem Biophys Res Commun 279:500–504

    Article  CAS  Google Scholar 

  • Qin AL, Zhou XQ, Zhang WQ et al (2004) Characteriation an enrichment of hepatic progenitor cells in adult liver. World J Gastroenterol 10:1480–1486

    CAS  Google Scholar 

  • Rosenberg IL, Russel CW, Giles GR (1978) Cell viability studies on the exfoliated colonic cancer cell. British J Sur 65:188–190

    Article  CAS  Google Scholar 

  • Sakai Y, Jiang J, Kojima N et al (2002) Enhanced in vitro maturation of fetal mouse liver cells with oncostatin M, nicotinamide, and dimethyl sulfoxide. Cell Transplant 11:435–441

    CAS  Google Scholar 

  • Seo MJ, Suh SY, Bae YC et al (2005) Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo. Biochem Biophys Res Commun 328:258–264

    Article  CAS  Google Scholar 

  • Talens R, Bonora A, Jover R et al (2006) Hepatogenic differentiation of human mesenchymal stem cells from adipose tissue in comparison with bone marrowmesenchymal stem cells. World J Gastroenterol 12:5834–5845

    Google Scholar 

  • Trey C, Davidson CS (1970) The management of fulminant hepatic failure. Prog Liver Dis 3:282–298

    CAS  Google Scholar 

  • Visconti RT, Bonora A, Jover R et al (2006) Hepatogenic differentiation of human mesenchymal stem cells from adipose tissue in comparison with bone marrow mesenchymal stem cells. World J Gastroenterol 28:5834–5845

    Google Scholar 

  • Wagner JE, Kernan NA, Steinbuch M et al (1995) Allogeneic sibling umbilical-cord-blood transplantation in children with malignant and non-malignant disease. Lancet 346:214–219

    Article  CAS  Google Scholar 

  • Wang X, Ge S, McNamara G et al (2003) Albumin-expressing hepatocyte-like cells develop in the livers of immune-deficient mice that received transplants of highly purified human hematopoietic stem cells. Blood 101:4201–4208

    Article  CAS  Google Scholar 

  • Wang PP, Wang JH, Yan P et al (2004) Expression of hepatocyte-like phenotypes in bone marrow stromal cells after HGF induction. Biochem Biophys Res Commun 320:712–716

    Article  CAS  Google Scholar 

  • Wells JM, Melton DA (2000) Early mouse endoderm is patterned by soluble factors from adjacent germ layers. Development 127:1563–1572

    CAS  Google Scholar 

  • Yu S, Li C, Xin-Guo H et al (2007) Differentiation of bone marrow-derived mesenchymal stem cell from diabetic patients into insulin-producing cells in vitro. Chinese Med J 120:771–776

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Zoology, University of Madras, Guindy Campus, Chennai, 600 025, Tamilnadu, India

    S. Sellamuthu, G. Babu, D. Dinesh, D. Prabhu & C. Arulvasu

  2. Department of Animal Health and Management, Alagappa University, Karaikudi, 03, Tamilnadu, India

    R. Manikandan

  3. Department of Biotechnology, School of Chemical and Biotechnology, Sastra University , Thanjavur, 613 401, Tamilnadu, India

    R. Thiagarajan

Authors
  1. S. Sellamuthu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R. Manikandan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R. Thiagarajan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. Babu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. D. Dinesh
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. D. Prabhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. C. Arulvasu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to C. Arulvasu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sellamuthu, S., Manikandan, R., Thiagarajan, R. et al. In vitro trans-differentiation of human umbilical cord derived hematopoietic stem cells into hepatocyte like cells using combination of growth factors for cell based therapy. Cytotechnology 63, 259–268 (2011). https://doi.org/10.1007/s10616-011-9337-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-011-9337-x

Keywords

Search

Navigation