title: In vitro generation of type-II pneumocytes can be initiated in human CD34(+) stem cells.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Stem Cell Laboratory, Department of Biotechnology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, 517507, India.
authors.forename: Lokanathan
authors.initials: L
authors.lastname: Srikanth
authors.affiliation: Stem Cell Laboratory, Department of Biotechnology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, 517507, India.
authors.forename: Katari
authors.initials: K
authors.lastname: Venkatesh
authors.affiliation: Stem Cell Laboratory, Department of Biotechnology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, 517507, India.
authors.forename: Manne Mudhu
authors.initials: MM
authors.lastname: Sunitha
authors.affiliation: Stem Cell Laboratory, Department of Biotechnology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, 517507, India.
authors.forename: Pasupuleti Santhosh
authors.initials: PS
authors.lastname: Kumar
authors.affiliation: Department of Hematology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, India.
authors.forename: Chodimella
authors.initials: C
authors.lastname: Chandrasekhar
authors.affiliation: Department of Neurology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, India.
authors.forename: Bhuma
authors.initials: B
authors.lastname: Vengamma
authors.affiliation: Stem Cell Laboratory, Department of Biotechnology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, 517507, India. sarmasvims@gmail.com.
authors.forename: Potukuchi Venkata Gurunadha Krishna
authors.initials: PV
authors.lastname: Sarma
mesh_terms: D056809:Alveolar Epithelial Cells / Q000502:physiology*; D018952:Antigens, CD34 / Q000032:analysis*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry; D005434:Flow Cytometry; D006801:Humans; D013234:Stem Cells / Q000737:chemistry* / Q000502:physiology*
chemical_list: D018952:Antigens, CD34; D003470:Culture Media
keywords: Air–liquid interface culture; CD34+ stem cells; Lysozyme; Real-time PCR; Stem cells; Surfactant proteins; Type-II pneumocytes