Ying Sun1,2, Yan Wang2, Zongjin Li1, Zhikun Guo1,2. 1. Henan Key Laboratory of Medical Tissue Regeneration, Xinxiang Medical University, Xinxiang, China. 2. Department of Cardiology, Zhengzhou Seventh People's Hospital, Zhengzhou, China.
Abstract
OBJECTIVE: The aim of the present study is to isolate and analyze the characterization of pericardial fluid cells (PFCs) from rat and provides a morphological basis for the basic research and clinical application of PFCs. METHODS: After aseptic thoracotomy was performed, normal saline was injected into the pericardial cavity of 50 adult Sprague-Dawley rats. The mixture of diluted pericardial fluid was extracted, centrifuged, and cultured. The cell morphology of different generations in the pericardial fluid was observed on an inverted microscope. The expression levels of CD44, CD29, CD90, and pan-hematopoietic marker CD45 were analyzed via flow cytometry. The third-generation cells were used for osteogenic, adipogenic, and cardiac differentiation. RESULTS: PFCs were successfully isolated and subcultured. PFCs were predominantly circular in shape after 24 h of culture. Following subculture for 3 days, the cells demonstrated a spindle shape. The rat pericardial fluid contains cell populations with uniform morphology, good growth state, and strong proliferation ability. Flow cytometry results showed that CD29 (100%) and CD90 (99.3%) were positively expressed, whereas CD45 (0.30%) and CD44 (0.48%) were negatively expressed. The PFCs could differentiate into osteoblasts and adipocytes after being induced. Cardiac differentiation was also confirmed by cardiac troponin T (cTnT) and α-sarcomeric actin (α-SA) staining. CONCLUSION: This study revealed that a subpopulation of cells was isolated from pericardial fluid, which exhibited progenitor cell features and multiple differentiation potency. PFCs could serve as an alternative cell source for myocardial tissue repair, engineering, and reconstruction.
OBJECTIVE: The aim of the present study is to isolate and analyze the characterization of pericardial fluid cells (PFCs) from rat and provides a morphological basis for the basic research and clinical application of PFCs. METHODS: After aseptic thoracotomy was performed, normal saline was injected into the pericardial cavity of 50 adult Sprague-Dawley rats. The mixture of diluted pericardial fluid was extracted, centrifuged, and cultured. The cell morphology of different generations in the pericardial fluid was observed on an inverted microscope. The expression levels of CD44, CD29, CD90, and pan-hematopoietic marker CD45 were analyzed via flow cytometry. The third-generation cells were used for osteogenic, adipogenic, and cardiac differentiation. RESULTS: PFCs were successfully isolated and subcultured. PFCs were predominantly circular in shape after 24 h of culture. Following subculture for 3 days, the cells demonstrated a spindle shape. The rat pericardial fluid contains cell populations with uniform morphology, good growth state, and strong proliferation ability. Flow cytometry results showed that CD29 (100%) and CD90 (99.3%) were positively expressed, whereas CD45 (0.30%) and CD44 (0.48%) were negatively expressed. The PFCs could differentiate into osteoblasts and adipocytes after being induced. Cardiac differentiation was also confirmed by cardiac troponin T (cTnT) and α-sarcomeric actin (α-SA) staining. CONCLUSION: This study revealed that a subpopulation of cells was isolated from pericardial fluid, which exhibited progenitor cell features and multiple differentiation potency. PFCs could serve as an alternative cell source for myocardial tissue repair, engineering, and reconstruction.
Authors: Elena A Jones; Anne English; Karen Henshaw; Sally E Kinsey; Alex F Markham; Paul Emery; Dennis McGonagle Journal: Arthritis Rheum Date: 2004-03