title: In vitro culture and characterization of enteric neural precursor cells from human gut biopsy specimens using polymer scaffold.
abstract: In vitro expansion and characterization of neural precursor cells from human gut biopsy specimens with or without Hirschsprung's disease using a novel thermoreversible gelation polymer (TGP) is reported aiming at a possible future treatment. Gut biopsy samples were obtained from five patients undergoing gut resection for Hirschsprung's disease (n = 1) or gastrointestinal disorders (n = 4). Cells isolated from the smooth muscle layer and the myenteric plexus were cultured in two groups for 18 to 28 days; Group I: conventional culture as earlier reported and Group II: using TGP scaffold. Neurosphere like bodies (NLBs) were observed in the cultures between 8th to 12th day and H & E staining was positive for neural cells in both groups including aganglionic gut portion from the Hirschsprung's disease patient. Immunohistochemistry using S-100 and neuron specific enolase (NSE) was positive in both groups but the TGP group (Group II) showed more number of cells with intense cytoplasmic granular positivity for both NSE and S-100 compared to Group I. TGP supports the in vitro expansion of human gut derived neuronal cells with seemingly better quality NLBs. Animal Studies can be tried to validate their functional outcome by transplanting the NLBs with TGP scaffolds to see whether this can enhance the outcome of cell based therapies for Hirschsprung's disease.
authors.affiliation: Department of Paediatric Surgery, Madras Medical College, E.V.R Periyar Salai, Park Town, Chennai, India; ; Department of Paediatric Surgery, Government Kasturba Gandhi Hospital, Triplicane, Chennai, India;
authors.forename: Janardhanam
authors.initials: J
authors.lastname: Krishnamohan
authors.affiliation: Department of Paediatric Surgery, Institute of Child Health & Hospital for Children, Egmore, Chennai, India;
authors.forename: Venugopal S
authors.initials: VS
authors.lastname: Senthilnathan
authors.affiliation: The Mary-Yoshio Translational Hexagon (MYTH), Nichi-In Centre for Regenerative Medicine (NCRM), Nungambakkam, Chennai, India;
authors.forename: Tirunelveli Muthiah
authors.initials: TM
authors.lastname: Vaikundaraman
authors.affiliation: The Fujio-Eiji Academic Terrain (FEAT), Nichi-In Centre for Regenerative Medicine (NCRM), Nungambakkam, Chennai, India;
authors.forename: Thangavelu
authors.initials: T
authors.lastname: Srinivasan
authors.affiliation: Ruma Biotherapy Research Centre, Alwarpet, Chennai, India;
authors.forename: Madasamy
authors.initials: M
authors.lastname: Balamurugan
authors.affiliation: Yamanashi University- School of Medicine, Chuo, Yamanashi, Japan;
authors.forename: Masaru
authors.initials: M
authors.lastname: Iwasaki
authors.affiliation: The Fujio-Eiji Academic Terrain (FEAT), Nichi-In Centre for Regenerative Medicine (NCRM), Nungambakkam, Chennai, India; ; Hope Foundation (Trust), Choolaimedu, Chennai, India.
authors.forename: Senthilkumar
authors.initials: S
authors.lastname: Preethy
authors.affiliation: The Mary-Yoshio Translational Hexagon (MYTH), Nichi-In Centre for Regenerative Medicine (NCRM), Nungambakkam, Chennai, India; ; Yamanashi University- School of Medicine, Chuo, Yamanashi, Japan;
authors.forename: Samuel Jk
authors.initials: SJ
authors.lastname: Abraham
keywords: Enteric neural precursor cells; Hirschsprung's disease; thermoreversible gelation polymer (TGP)