The Isolation and Characterization of Muscle Derived Stem Cells from Gastrocnemius Muscle of Rats Using the Modified Preplate Method / 대한비뇨기과학회지
Korean Journal of Urology
; : 1279-1284, 2004.
Article
in Korean
| WPRIM (Western Pacific)
| ID: wpr-144322
Responsible library:
WPRO
ABSTRACT
PURPOSE:
This study attempted to characterize the muscle derived stem cells isolated from the primary cultured skeletal muscle of the rat gastrocnemius muscle; in addition, we modified the preplate method and then compared this to the original preplate method. MATERIALS ANDMETHODS:
The hind limbs (gastrocnemius muscles) were removed from a 3-6 week olds SD-rat and the bone was dissected away. The muscle mass was finely minced and chopped using razor blades. In an original preplate method, the cells were dissociated using a triple enzyme mixture (collagenase XI, dipase and trypsin) for 1 hour at 37degreesC. The muscle cell extract was preplated on culture flasks as described by Dr. Qu (Qu et al., 1998). The pp1-pp4 cells were referred to as the early plate (EP) cells, and the pp5-pp6 cells were referred to as the late plate (LP) cells. When we modified the preplate method, the pp1-pp2 cells were called the early plate (EP) cells and the pp3-pp4 cells were called to late plate (LP) cells. The phenotypical characteristics of EP and LP cells were compared by immunostaining and FACS.RESULTS:
In the original preplate methods, the early plate (EP) cells were mixed with myogenic cells (mostly fibroblasts, 90% desmin + cells). Yet in the modified preplate method, the muscle derived stem cells were determined to be CD34 (+ or -), CD45- and desmin- cells by immunohistochemical staining and FACS.CONCLUSIONS:
In original methods, the LP cells exhibited stem cell properties (CD34+, less than 30%), and they were not from a hematogeous origin (CD45-), but rather, they were from a myogenic origin (desmin+). Yet in the modified preplate method, we purified the LP cells much earlier than the original method. The LP cells displayed CD34+(more than 50%), and CD45-; thus, we isolated more primitive (desmin-) cells.
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Stem Cells
/
Muscle, Skeletal
/
Muscle Cells
/
Desmin
/
Extremities
/
Fibroblasts
Limits:
Animals
Language:
Korean
Journal:
Korean Journal of Urology
Year:
2004
Document type:
Article