Isolation and characterization of porcine adipose tissue-derived adult stem cells.
BACKGROUND
Stem cell characteristics such as self-renewal, differentiation and expression of CD34 and CD44 stem cell markers have not been identified in porcine adipose tissue-derived adult stem (ADAS) cells. The objective of this study was to develop a protocol for the isolation and culture of porcine adipose tissue-derived cells and to determine stem cell-like characteristics.


METHODS
Primary cultures were established and cell cultures were maintained. Cloning capacity was determined using a ring cloning procedure. Primary cultures and clones were differentiated and stained for multiple differentiated phenotypes. CD34 and CD44 messenger ribonucleic acid (mRNA) was isolated and reverse transcriptase polymerase chain reaction was used to compare expression profiles.


RESULTS
An average of 2,700,000 nucleated cells/ml was isolated; 26% were adherent, and cells completed a cell cycle approximately every 3.3 days. Ring cloning identified 19 colonies. Primary cultures and clones were determined to differentiate along osteogenic, adipogenic and chondrogenic tissue lineages. The mRNA expression profiles showed CD34 expression was higher for undifferentiated ADAS cells versus differentiated cell types and the CD34 expression level was lower than that of CD44 among differentiated cells.


CONCLUSIONS
Improved culture conditions and defined cellular characteristics of these porcine ADAS cells have been identified. Porcine ADAS can self-renew, can differentiate into multiple tissue lineages and they express CD34.
Williams
Kellie J
KJ
School of Animal Sciences, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA.
Picou
Alicia A
AA
Kish
Sharon L
SL
Giraldo
Angelica M
AM
Godke
Robert A
RA
Bondioli
Kenneth R
KR
D000273:Adipose Tissue / Q000166:cytology*; D053687:Adult Stem Cells / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics; D002454:Cell Differentiation; D002478:Cells, Cultured; D005260:Female; D018960:Hyaluronan Receptors / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine
D018952:Antigens, CD34; D018960:Hyaluronan Receptors