Home » Laboratories » F1a. Cell Culture Lab

Purpose:

The Cell Culture Lab is an essential component of a Translational Medicine platform. Within the Department of Gastroenterology and Hepatology of the CEMT, a Molecular and Cellular Biology Laboratory was designed according to the biosecurity standards  for a BSL 2 level.

Inside this lab we can create cell cultures (cell lines and primary cells), primary cell isolation activities for in vitro studies (eg hepatocyte isolation from liver tissue fragments, digestive epithelial cell isolation), ectopic gene expression studies , customized cell lines can be generated using pluripotent cells induced for personalized genetic and pharmacological studies.

Laboratory design

  • 3 Laboratories for molecular and cell biology experiments
  • Image acquisition and image processing area
  • Biological samples storage room (-80C)
  • Four degree room (4C)
  • Area designated to processing of waste material resulted from research activities
  • Area designated to research planning and documentation

Research Activities

  • isolation of human hepatocytes and digestive epithelial cells and research of new in vitro cultivation techniques aiming at functional preservation
  • hepatocyte / hepatic proliferative hepatic transdifferentiation studies by ectopic expression of transcription factors
  • obtaining human hepatocyte-like cells customized by cellular reprogramming techniques with potential utility in personalized pharmacology studies and regenerative medicine;
  • the study of epithelial-mesenchymal transition phenomena and hepatocyte reprogramming under conditions of chronic liver injury.
  • the role of ectopic expression of transcription factors as a method of regenerative gene therapy in chronic diseases and antitumor therapy
  • Generation of in vitro cellular modeling models using new genomic editing techniques

The BSL 2.1 Laboratory is dedicated to procedures of isolation of primary cell cultures from tissue fragments, culture expansion and generating cell banks consisting of both primary cells and cell lines for in Vitro research.

BSL 2.2 Laboratory

The BSL 2.2 Laboratoryis dedicated to procedures that involve genetic modification of cells (viral transduction, plasmid transfections) and generatingin vitrostudy models, using new genome editing techniques.

Biosecurity level: BSL2 +

BSL2.3 Laboratory

The BSL2.3 laboratory  is designed for the generation of plasmid vectors for ectopic expression of genes of interest: PCR amplification of the genes, enzymatic digestion of DNA using specific restriction enzymes, agarose gel electrophoresis of nucleic acids, cloning of nucleic acid sequences with the generation of specific plasmids and transformation of competent bacteria for plasmid production, bacterial cultures in liquid and solid medium, using both static incubator and shaker and agitator, isolation of plasmids from bacterial cultures.

Image acquisition and image processing:
  • this area is dedicated to image acquisition from cell cultures using the inverted microscope with ability to generate phase contrast, DIC and fluorescence microscopy images.

Long-term storage room for biological samples: stores in low temperature conditions biological samples (tissue, biological fluids, cell cultures, genetically modified cell lines, plasmid or viral vectors) in a freezer with very low temperatures (-80°C), as well as in a storage container with a reservoir of liquid nitrogen (-196°C).

 

Waste Processing Area: where the waste resulted from research activity is collected and autoclaved before being removed from the laboratory.

Working space for documentation and preparation of research materials: 3 working stations for the researchers that work in this laboratory, software for gene sequence editing, Vector NTI Advance.

Research Projects currently implemented in the Laboratory:
  • DIACURE: The project is aimed at transdifferentiationof hepatic cells to beta pancreatic cells, by ectopic expression of specific transcription factors. The project opens new perspectives for autologous cell therapy in diabetes mellitus.
  • APOHEPI: The main objective of the research project is to study the effects of immunosuppressive therapy used in clinical practice of liver transplantation on apoptosis of hepatocytes and stellate cells, in relationship with specific gene polymorphisms.
  • PANCNGS: In vitro studies of pancreatic carcinogenesis using transgenic cell lines expressing key genetic mediators of the carcinogenic process.
  • THERRES: In vitro studies of gastric carcinogenesis using transgenic cell lines obtained by CRISPR CAS9 technology

Members

Razvan Iacob
Senior Physician in Gastroenterology, , MD, PhD

Speranta Iacob
Senior Physician in Gastroenterology

Mihaela Ecaterina Uță
Research Scientist

INNOVATIVE RESEARCH
OF EUROPEAN LEVEL

CENTER OF EXCELLENCE IN TRANSLATIONAL
MEDICINE

The strategic goal is to create research cores with advanced technological facilities in the form of research platforms by associating preclinical research laboratories and clinical research centers within the Fundeni Clinical Institute in a unique entity.

The newly created structure provides logistical support for biomedical research through the coordination of studies, quality management, data monitoring, dissemination of good clinical practice, database management and data analysis. Infrastructure is also dedicated to the production of vectors for gene and cellular therapy, as well as for the acquisition of new agents with therapeutic potential in digestive affection.

Subscribe to
CEMT Newsletter

For more informations please subscribe to our newsletter.
Fields with * are required.