283 In Vitro Cell. Dev. Biol.—Animal 42:283–286, November/December 2006  2006 Society for In Vitro Biology 1071-2690/06 $18.00+0.00 HUMAN UMBILICAL CORD BLOOD SERUM PROMOTES GROWTH, PROLIFERATION, AS WELL AS DIFFERENTIATION OF HUMAN BONE MARROW–DERIVED PROGENITOR CELLS SMRUTI M. PHADNIS, MUGDHA V. JOGLEKAR, VIJAYALAKSHMI VENKATESHAN, SURENDRA M. GHASKADBI, ANANDWARDHAN A. HARDIKAR, AND RAMESH R. BHONDE 1 Division of Animal Sciences, Agharkar Research Institute, Pune, 41004 India (S. M. P., S. G.), Tissue Engineering and Banking Laboratory (M. V. J., R. R. B.) and Stem Cell and Diabetes Section, National Center for Cell Science, Pune, 41107 India (A. A. H.) and Department of Biochemistry, National Institute of Nutrition, Jamia Osmania, Hyderabad, 500007 India (V. V.) (Received 15 March 2006; accepted 20 March 2006) SUMMARY Fetal calf serum (FCS) is conventionally used for animal cell cultures due to its inherent growth-promoting activities. However animal welfare issues and stringent requirements for human transplantation studies demand a suitable alternative for FCS. With this view, we studied the effect of FCS, human AB serum (ABS), and human umbilical cord blood serum (UCBS) on murine islets of Langerhans and human bone marrow–derived mesenchymal-like cells (hBMCs). We found that there was no difference in morphology and functionality of mouse islets cultured in any of these three different serum supplements as indicated by insulin immunostaining. A comparative analysis of hBMCs maintained in each of these three different serum supplements demonstrated that UCBS supplemented media better supported proliferation of hBMCs. Moreover, a modification of adipogenic differentiation protocol using UCBS indicates that it can be used as a supplement to support differentiation of hBMCs into adipocytes. Our results demonstrate that UCBS not only is suitable for mainte- nance of murine pancreatic islets, but also supports attachment, propagation, and differentiation of hBMCs in vitro. We conclude that UCBS can serve as a better serum supplement for growth, maintenance, and differentiation of hBMCs, making it a more suitable supplement in cell systems that have therapeutic potential in human transplantation programs. Key words: umbilical cord blood serum; fetal calf serum; islets; bone marrow; serum supplement. In vitro culture systems available for various cell types form an ideal model for substitution of animal experiments in majority of cases (Sasse et al., 2000). In most of these culture systems, fetal bovine serum/fetal calf serum (FCS) is an essential component in the growth and maintenance of cells. It is a complex mixture of low and high molecular weight biomolecules with a variety of growth- promoting and growth-inhibiting properties. Low toxicity and growth-promoting properties of FCS make it a popular supplement for in vitro cultivation of mammalian cells. However, animal sera are a potential source of microbiological contaminants, notably my- coplasma, bovine viruses, and other pathogens, and agents that can cause bovine spongiform encephalopathy or Creutzfeldt-Jakob dis- ease (Klein and Dumble, 1993). Suppliers use a series of tech- niques, including filtration, irradiation, and/or heat-inactivation, to reduce any microbial contamination. As a result of these issues, cell and organ transplantation programs do not recommend the use of calf/bovine serum supplement (Koivisto et al., 2004). Moreover, clinical usage of FCS also involves local inflammation, antibody production against xenoproteins, and nonengraftment due to xeno- genic immune responses (Stute et al., 2004). All these factors raise 1 To whom correspondence should be addressed at E-mail: rrbhonde@ nccs.res.in the question of biosafety involved in the use of FCS in cell systems that carry therapeutic potential. Alternatives to FCS include use of goat serum (Paranjape, 2004), bovine colostrums (Capiaumont et al., 1996), bovine ocular fluid in combination with sheep’s defibrinated plasma, and human AB se- rum ([ABS]; Filipic et al., 2002). Earlier attempts to replace FCS by goat serum in cell culture, especially hybridoma cells, met with little success due to toxicity of goat serum. Addition of soybean lipid mixture to goat serum was proposed to make it compatible with FCS; however, long-term cultures demonstrated higher mortal- ity with goat serum (Deshpande et al., 2000). Other than these supplements, it also is possible in some cases to use a fully chem- ically defined media with combination of appropriate hormones and growth factors. A compilation of commercially available serum-free media is available on http://www.focusonalternatives.org.uk. Human mesenchymal stem cells (hMSCs) grown in autologous serum were seen to expand rapidly with stable gene expression in absence of growth factors, whereas FCS was reported to induce more differ- entiated and less stable transcript profile (Shahdadfar, 2005). Hu- man AB serum also has been used for hMSCs with no detectable alterations in proliferation or differentiation potential in vitro and in vivo (Yamaguchi, 2002; Kassem, 2004). Survival of granulocyte monocyte—colony-forming cells from patients with chronic granu- locytic leukemia in liquid culture has been shown to be dependent