Utility of human placental alkaline phosphatase as a genetic marker for cell tracking in bone and cartilage.
It was the aim of the current study to evaluate the utility of human placental alkaline phosphatase (hPLAP) as a genetic marker for cell tracking in bone and cartilage, using transgenic Fischer 344 rats expressing hPLAP under the control of the ubiquitous R26 promoter [F344-Tg(R26-hPLAP)]. hPLAP enzyme activity was retained during paraffin and methylmethacrylate (MMA) embedding, and was best preserved using 40% ethanol as fixative. Endogenous alkaline phosphatase activity could be completely blocked by heat inactivation in paraffin and MMA sections, allowing histochemical detection of hPLAP in the complete absence of background staining. In addition, sensitive detection of hPLAP was also possible using immunohistochemistry. F344-Tg(R26-hPLAP) rats demonstrated ubiquitous expression of hPLAP in hematopoietic bone marrow cells and stromal cells such as osteoblasts, osteocytes, and chondrocytes. Osteoclasts only weakly expressed hPLAP. In conclusion, hPLAP provides superb detection quality in paraffin and plastic sections, and constitutes an excellent genetic marker for cell tracking in hard and soft tissues.
Unger
Nina J
NJ
Institute of Physiology, Physiological Chemistry and Animal Nutrition, Ludwig Maximilians University, Munich, 80539, Germany.
Odörfer
Kathrin I
KI
Weber
Karin
K
Sandgren
Eric P
EP
Erben
Reinhold G
RG
D000469:Alkaline Phosphatase; D000818:Animals; D030801:Animals, Genetically Modified; D001854:Bone Marrow Cells / Q000166:cytology / Q000201:enzymology; D001842:Bone and Bones / Q000166:cytology / Q000201:enzymology*; D002356:Cartilage / Q000166:cytology / Q000201:enzymology*; D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage / Q000502:physiology*; D019902:Chondrocytes / Q000166:cytology / Q000201:enzymology; D005260:Female; D058851:GPI-Linked Proteins; D005819:Genetic Markers; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008297:Male; D010006:Osteoblasts / Q000166:cytology / Q000201:enzymology; D010010:Osteoclasts / Q000166:cytology / Q000201:enzymology; D010011:Osteocytes / Q000166:cytology / Q000201:enzymology; D051381:Rats; D011916:Rats, Inbred F344; D017154:Stromal Cells / Q000166:cytology / Q000201:enzymology
D058851:GPI-Linked Proteins; D005819:Genetic Markers; D007527:Isoenzymes; D000469:Alkaline Phosphatase; C019839:alkaline phosphatase, placental