The document discusses mesenchymal stem cells (MSCs) and their growth factors. It finds that MSCs reach senescence after a limited number of cell divisions due to induction of the p16 protein. While overexpression of telomerase and cell cycle regulatory genes can prolong the lifespan of MSCs, the key inducers of p16 expression are platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF). In particular, PDGF subunit B is responsible for p16 expression in MSCs. The type of growth factor stimulus affects p16 expression levels in the cells.
This document discusses research on using mesenchymal stem cells and biomaterials for regenerative medicine applications. Specifically, it examines using marrow stromal cells and engineered scaffolds to generate bone and cartilage tissues. It describes experiments showing marrow stromal cells can form bone in vivo and discusses strategies for controlling the shape of regenerated tissues using cell-seeded biodegradable polymer sheets and scaffolds.
This document discusses research on marrow stromal cells and their potential applications for regenerative medicine and tissue engineering. Key points include:
1. Marrow stromal cells reside in the bone marrow microenvironment and can differentiate into various cell types, including bone, cartilage, fat, and muscle cells.
2. Studies show marrow stromal cells implanted in rodents generate new bone tissue in vivo.
3. The document proposes strategies for controlling the shape of new bone tissue using biodegradable polymer scaffolds seeded with osteoblasts or chondrocytes.
4. Methods are presented for selecting progenitor cells using cell surface markers and gene regulatory elements to target cell differentiation for tissue engineering applications
8. Human Marrow Stromal Cells
reach senescence after a limited number of cell division
H4-1H4-1
H4-1H4-1
慶應義塾大学医学部慶應義塾大学医学部
血液内科血液内科
木崎 昌弘 木崎 昌弘
福地 由美 福地 由美
H4-1, senescenceH4-1, senescence
9. Prolongation of the life span of human marrow stromal cell
by transferring Bmi-1, E6, E7, and TERT
The cells with the extended life span retained the multipotency
into osteocytes, chondrocytes, adipocytes, and myocytes in vitro.
12. 1 week 3 weeks
Recording of action potentials
from spontaneously beating cells
Alexa568
Human marrow stromal cells
with the extended life span
can differentiate into cardiomyocytes
Disorganized
Regular and
stabilized
Recording
microelectrode
20. EPC-100 (2W) Patch clamp Fast Na current was observed in 1/10 cell
EPC-214 (2W) Patch clamp Fast Na current was observed in 1/5 cell
EPC
pipette
Patch clamp from post-FACS EPCs
GFP を陽性細胞 FACS にて 、その から抽出 細胞 patch clamp を った行
22. Non
fusion
EPC-100 AEPC-214
Hoechest EGFP h-cardiac troponin-I Merge
Hoechest EGFP h-cardiac troponin-I Merge
B C D E
F G H I
Feeder cardiomyocyte
Collagen membraneculture dish
EPC
44. SPIO 効果 ; T2 短縮(信号低下)
T2 強調 SPIO 造影 T2 強調
•正常肝細胞は鉄コロイド
を貪食するので信号が低下
する•貪食能のない病変は信号
が低下しないので白く浮き
出て見える
Labeling
Fe
Fe
Fe
Fe
Fe Fe
Fe Fe
Fe
Fe
Fe Fe
Fe
Fe
Fe
Fe
FeFe
Fe Fe
Fe
Fe
Fe
Fe
Fe
Phagocytosis
or
Pinocytosis
SPIO labeled cell
SPIO
52. BB(+) Vim(-)
= mouse
macrophage
BB(-) Vim(+)
=survived
human cells
BB(+) Vim(+)
=survived
human cells
Berlin Blue stain + Vimentin stain 4 weeks
53. Time course
after transplantation
移植細胞内に SPIO 存在
移植細胞からの排出
マクロファージの遊走開始
マクロファージによる鉄貪飲
移植細胞の形態変化
0
W
1
W
2
W
4
W
8
W
マクロファージのアポトーシス?
鉄の消失(網内系への取り込み?)
The fate of implanted bone marrow stromal cells
移植細胞の局在確認