Cytotoxicity of rat marrow stromal cells against malignant glioma cells.
OBJECTIVE
Marrow stromal cells (MSCs) have been shown to have the capacity of orthodox and unorthodox plasticity. In this study, the authors tried to access in vitro cytotoxicity of MSCs from rat and also to differentiate MSCs into immune effector cell.


METHODS
Rat MSCs (rMSCs) were isolated by standard methodology and were activated by interleukin-2 (IL-2), interleukin-15 (IL-15), granulocyte macrophage colony stimulating factor, and combinations, which were effector cells. Cytotoxicity of rMSCs and activated rMSCs against the target cells (9L rat glioma cell line) was estimated using visual survival cell assay. Phenotypes of these various activated cells were determined using flow cytometry. The secreted protein from effector cells was estimated by enzyme-linked immunosorbent assay. The expression of immune response-related genes in activated cells was measured.


RESULTS
There was a significant cytotoxicity of rMSCs activated with various cytokine combinations. After various cytokine activations of rMSCs, the population of immune effector cells (CD8, CD161a) and immune reaction-related proteins (IL-4, gamma-INF) might increase. Apoptosis may be one of the lysis mechanisms of target cells by activated rMSCs. The contributing genes could be gamma-INF, FasL, and perforin.


CONCLUSIONS
This study suggests that rMSC may be used as adoptive transfer therapy in patients suffering from malignant brain tumor, but we have to investigate orthotopic animal study for the proper translation.
Kang
Seok-Gu
SG
Department of Neurosurgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Gangnam-gu, Seoul, Republic of Korea.
Jeun
Sin Soo
SS
Lim
Jung Yeon
JY
Yoo
Do Sung
DS
Huh
Pil Woo
PW
Cho
Kyung Souk
KS
Kim
Dal Soo
DS
Shin
Hyung-Jin
HJ
Kim
Jong Hyun
JH
Kim
Moon Chan
MC
Kang
Joon Ki
JK
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015152:Blotting, Northern / Q000379:methods; D015139:Blotting, Southern / Q000379:methods; D001854:Bone Marrow Cells / Q000187:drug effects / Q000502:physiology*; D002452:Cell Count / Q000379:methods; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D017077:Culture Media, Conditioned / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology*; D004338:Drug Combinations; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D053222:Fas Ligand Protein; D005434:Flow Cytometry / Q000379:methods; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D005910:Glioma / Q000628:therapy*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D020287:In Situ Nick-End Labeling / Q000379:methods; D015847:Interleukin-4 / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D017154:Stromal Cells / Q000502:physiology*; D014162:Transfection / Q000379:methods; D048069:Tumor Necrosis Factors / Q000235:genetics / Q000378:metabolism
D015703:Antigens, CD; D017077:Culture Media, Conditioned; D016207:Cytokines; D004338:Drug Combinations; D053222:Fas Ligand Protein; C505662:Faslg protein, rat; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D048069:Tumor Necrosis Factors; D049452:Green Fluorescent Proteins; D015847:Interleukin-4