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Intractable Rare Dis Res. 2013 Aug;2(3):98-102. doi: 10.5582/irdr.2013.v2.3.98.

In vitro culture and characterization of enteric neural precursor cells from human gut biopsy specimens using polymer scaffold.

Author information

1
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;
2
Department of Paediatric Surgery, Institute of Child Health & Hospital for Children, Egmore, Chennai, India;
3
The Mary-Yoshio Translational Hexagon (MYTH), Nichi-In Centre for Regenerative Medicine (NCRM), Nungambakkam, Chennai, India;
4
The Fujio-Eiji Academic Terrain (FEAT), Nichi-In Centre for Regenerative Medicine (NCRM), Nungambakkam, Chennai, India;
5
Ruma Biotherapy Research Centre, Alwarpet, Chennai, India;
6
Yamanashi University- School of Medicine, Chuo, Yamanashi, Japan;
7
The Fujio-Eiji Academic Terrain (FEAT), Nichi-In Centre for Regenerative Medicine (NCRM), Nungambakkam, Chennai, India; ; Hope Foundation (Trust), Choolaimedu, Chennai, India.
8
The Mary-Yoshio Translational Hexagon (MYTH), Nichi-In Centre for Regenerative Medicine (NCRM), Nungambakkam, Chennai, India; ; Yamanashi University- School of Medicine, Chuo, Yamanashi, Japan;

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.

KEYWORDS:

Enteric neural precursor cells; Hirschsprung's disease; thermoreversible gelation polymer (TGP)

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