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Stem Cell Reviews and Reports

, Volume 7, Issue 4, pp 860–868 | Cite as

Human Serum is as Efficient as Fetal Bovine Serum in Supporting Proliferation and Differentiation of Human Multipotent Stromal (Mesenchymal) Stem Cells In Vitro and In Vivo

  • Abdullah Aldahmash
  • Mandana Haack-Sørensen
  • May Al-Nbaheen
  • Linda Harkness
  • Basem M. Abdallah
  • Moustapha KassemEmail author
Article

Abstract

Background

Human multipotent stromal (skeletal, mesenchymal) stem cells (hMSC) are employed in an increasing number of clinical trials for tissue regeneration of age-related degenerative diseases. However, routine use of fetal bovine sera (FBS) for their in vitro expansion is not optimal and may pose a health risk for patients.

Methods

We carried out a side-by-side comparison of the effects of allogenic pooled human serum (HuS) versus FBS on hMSC proliferation and differentiation in vitro and in vivo. As a model for hMSC, we employed telomerase-immortalized hMSC; hMSC-TERT cell line.

Results

hMSC-TERT exhibited similar morphology and size when cultured in HuS vs. FBS as assessed by light microscopy and FACS analysis. We did not observe any significant differences in growth rates of hMSC-TERT during short-term (10 days) and long-term (100 days) culture in media supplemented with HuS vs. FBS. hMSC-TERT or primary bone marrow derived hMSC induced to osteoblastic or adipocytic differentiation in the presence of HuS or FBS showed comparable levels of gene expression and protein production of osteoblastic markers (CBFA1/Runx2, alkaline phosphastase, collagen type I and osteocalcin) or adipocytic markers (PPAR-gamma2, lipoprotein lipase (LPL), aP2), respectively. In order to test for the functional capacity of hMSC-TERT that have been maintained in long-term cultures in the presence of HuS vs. FBS, the cells were mixed with hydroxyapatite/tricalcium phosphate (HA/TCP) and implanted subcutaneously in immune deficient mice. hMSC maintained in HuS vs. FBS formed comparable heterotopic bone.

Discussion

Human serum can support proliferation and differentiation of hMSC in vitro and can maintain their bone forming capacity in vivo. The use of human serum in cell cultures of hMSC intended for cell-based therapy is preferable.

Keywords

Human mesenchymal stem cells Stromal stem cells Fetal bovine serum Human serum Osteoblast differentiation Adipocyte differentiation 

Notes

Acknowledgement

The study was supported by grants from the Novo Nordisk Foundation, the Lundbeck foundation and grants from the Region of Southern Denmark and King Abdulaziz City for Science and Technology (09-BIO740-20).

Disclosure of Interest

The authors disclose no conflict of interest

Supplementary material

12015_2011_9274_Fig5_ESM.jpg (81 kb)
Supplementary Fig. 1

(a) Phase-contrast pictures of hMSC-TERT cultured in medium containing fetal bovine serum (FBS, 10%) or human serum (HuS, 5%).Magnification: 200×. (b) FACS analysis of size versus granularity of hMSC-TERT cells cultured in FBS (10%) or HuS (5%) (JPEG 80 kb)

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High resolution image (TIFF 2721 kb)
12015_2011_9274_Fig6_ESM.jpg (2.1 mb)
Supplementary Fig. 2

Cytochemical and immunocytochemical staining of hMSC-TERT cultured in osteoblast induction medium (induced) and control medium (control) in presence of fetal bovine serum FBS (10%) and human serum (HuS, 5%), magnification 100×. Alkaine phosphatase (AP), collagen type I, bone sialoprotein (BSP), osteopontin (OP), osteocalcin and alizarin R for mineralized matrix (AR-S). Magnification 200× (JPEG 2155 kb)

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High resolution image (TIFF 8460 kb)
12015_2011_9274_Fig7_ESM.jpg (1.9 mb)
Supplementary Fig. 3

Formation of adipocytes by hMSC-TERT cells cultured in adipocyte induction medium (induced) and control medium (control) in presence of human serum (HuS, 5%) and fetal bovine serum (FBS, 10%) and visualized by Oil red O staining. Magnification: 200× (JPEG 1938 kb)

12015_2011_9274_MOESM3_ESM.tif (3.5 mb)
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12015_2011_9274_Fig8_ESM.jpg (104 kb)
Supplementary Fig. 4

Effect of HuS versus FBS on osteoblast and adipocyte differentiation of primary hMSC. Primary hMSC were isolated and cultured as described in M&M. Cells were induced to either osteoblast or adipocyte differentiation in the presence of 10% HuS or 10% FBS for 12 days. (a) Quantitative ALP activity after normalization to cell viability as described in M&M. (b) ALP staining after 12 days in osteoblast induced medium. (c) % adipocyte quantification. (d) Oil red O staining of fat droplets after 12 days in adipogenic induction medium. Cells cultured in 10% HuS or FBS without induction media were used as controls in each experiment. Values are represented as means ± SD of three independent experiments (JPEG 104 kb)

12015_2011_9274_MOESM4_ESM.tif (2.8 mb)
High resolution image (TIFF 2899 kb)

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Abdullah Aldahmash
    • 1
    • 2
  • Mandana Haack-Sørensen
    • 1
  • May Al-Nbaheen
    • 2
  • Linda Harkness
    • 1
  • Basem M. Abdallah
    • 1
  • Moustapha Kassem
    • 1
    • 2
    • 3
    Email author
  1. 1.Endocrine Research Laboratory (KMEB), Department of Endocrinology and MetabolismOdense University Hospital & University of Southern DenmarkOdenseDenmark
  2. 2.Stem Cell Unit, Department of Anatomy, College of MedicineKing Saud UniversityRiyadhSaudi Arabia
  3. 3.University Department of Endocrinology and Metabolism, University Hospital of OdenseOdense CDenmark

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