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In vitro generation of type-II pneumocytes can be initiated in human CD34+ stem cells

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Abstract

Objectives

Human CD34+ stem cells differentiated into type-II pneumocytes in Dulbecco’s modified Eagle medium (DMEM) having hydrocortisone, insulin, fibroblast growth factor (FGF), epidermal growth factor (EGF) and bovine serum albumin (BSA), expressing surfactant proteins-B (SP-B) and C (SP-C), alkaline phosphatase (ALP) and lysozyme.

Results

FACS-enumerated pure CD34+ cells, isolated from human peripheral blood, were cultured in DMEM and showed positive reaction with anti-human CD34 monoclonal antibodies in immunocytochemistry. These cells were cultured in DMEM having hydrocortisone, insulin, FGF, EGF and BSA (HIFEB-D) medium having an air–liquid interface. They differentiated into type-II pneumocytes with expression of SP-B and SP-C genes and disappearance of CD34 expression as assessed using real-time PCR. In reverse transcription-PCR amplicons showed 208 and 907 bp confirming SP-B and SP-C expressions. These cells expressed ALP with an activity of 1.05 ± 0.09 mM ml−1 min−1 and lysozyme that killed E. coli.

Conclusion

The successful differentiation of human CD34+ stem cells into type-II pneumocytes, and transplantation of such cells obtained from the patient’s stem cell could be the futuristic approach to regenerate diseased lung alveoli.

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Acknowledgments

We sincerely acknowledge Sri Venkateswara Institute of Medical Sciences (SVIMS University), India for providing funds to carry out the work. This paper forms a part of Ph.D. thesis work going to be submitted to SVIMS University, Tirupati, Andhra Pradesh, India.

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Authors and Affiliations

  1. Stem Cell Laboratory, Department of Biotechnology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, 517507, India

    Lokanathan Srikanth, Katari Venkatesh, Manne Mudhu Sunitha, Pasupuleti Santhosh Kumar & Potukuchi Venkata Gurunadha Krishna Sarma

  2. Department of Hematology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, India

    Chodimella Chandrasekhar

  3. Department of Neurology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, India

    Bhuma Vengamma

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  1. Lokanathan Srikanth
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  2. Katari Venkatesh
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  3. Manne Mudhu Sunitha
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  4. Pasupuleti Santhosh Kumar
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  5. Chodimella Chandrasekhar
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  6. Bhuma Vengamma
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  7. Potukuchi Venkata Gurunadha Krishna Sarma
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Corresponding author

Correspondence to Potukuchi Venkata Gurunadha Krishna Sarma.

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Srikanth, L., Venkatesh, K., Sunitha, M.M. et al. In vitro generation of type-II pneumocytes can be initiated in human CD34+ stem cells. Biotechnol Lett 38, 237–242 (2016). https://doi.org/10.1007/s10529-015-1974-2

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  • DOI: https://doi.org/10.1007/s10529-015-1974-2

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