Professional Documents
Culture Documents
Introduction
science:
- biomaterials,
- growth factors, &
- stem cells to repair failing organs
ADSCs stem cell source w/ therapeutic
applicability in diverse fields repair &
regeneration of acute & chronically damaged
tissues
ADSc = BMSC
cell surface marker
gene expression profile
differentiation potential
ADSc >> BMSC
obtained in large quantities @ low risk
yields far more stem cell than BM (5000:100-
1000)
Lipoaspirate obtained from PAL and manual (1) were filtered through a fine mesh stainless steel tea or coffee filter (=7 cm) to separate adipose tissue
fragments from tumescent, blood, or contaminants (2), and then wash the fat tissue fragments by soaking the filter in a sterile porcelain bowl filled with
Phosphate Buffer Saline (PBS 1x) pH 7.4 and stirring using a small tea spoon to remove the other contaminants (3). This washing step is repeated
several times. When fat tissue fragments appeared yellow and clean, then transferred into sterile 50 ml centrifuge tube (4). 0.075% collagenase type 1
(Sigma, USA) is added into the tube containing fat tissue fragments until tissue fragments : collagenase solution was 1:2 (f5). The tubes then
incubated at 37oC, 5% CO2 for 1 hour with agitation every 5 minutes. After 1 hour, the appeared floating yellow free lipid were removed (6 & 7) and
infranatant were poured into new centrifuge tube through nylon mesh filter and divided into sterile 15 ml centrifuge tube (8 & 9) and then centrifuge
1200 rpm for 10 minutes. After centrifugation step, the sedimented pellet were added lysis buffer if red blood cell still appeared (10 &11), then
centrifuge step repeated, obtained pellet (12) then resuspended in culture medium. The number of viable cell were counted using Hemacytometer.
The cells were cultured in 12-well plate with seeding number around 170.000 viable cells per well). The cultures cells were observed and medium
changed every 2 3 days.
Description
CANUL
A
MICROA
IRE
Lipoaspiraate
Dark
red
Bright
red
Isolation
processing
time/washing
process in the lab
>>
>>>
Sample after
washing process
with PBS several
times
Bright,
Yellowis
yelowwi h, still
sh
seems
reddish
Remark
Description
CANUL
A
MICROAIRE
Sample after
digestion with
Collagenase
No RBC
Still there is
RBC (RBC
still present
while filtering
infranatant
through
nylon mesh
filter)
Clearer
Remark
Day-2
Manual
Canula
Microair
e
Day-4
Day-6
Day-9
Patient ID
Age
Sex
BMI
Harvest site
24
55
23.76
25
39
29.1
26
55
25.45
27
32
29.1
Abdomen
28
41
41.24
Abdomen
29
42
28.85
Abdomen
Flank
Abdomen
Flank
Patient ID
Age
24
Sex
BMI
23.76
Harvest site
Flank
Patient
ID
24
55
25
29.1
Abdomen
25
25.45
Flank
26
27
29.1
Abdomen
27
28
41.24
Abdomen
28
41
29
42
28.85
Abdomen
29
300 ml
1,660,000
11 G
300 ml
2,460,000
4.0 mm
300 ml
900,000
11 G
300 ml
1,310,000
4.0 mm
100 ml
855,000
11 G
300 ml
1,920,000
1,883,333
4.0 mm
500 ml
11 G
<500 ml
5.00 mm
>300 ml
11 G
>300 ml
4.0 mm
300 ml
11 G
250 ml
PAL
Manual Canula
3.0 mm
PAL
Manual Canula
32
Cell yield
count
PAL
Manual Canula
55
Lipoaspirate
Volume
PAL
Manual Canula
39
Canula type
PAL
Manual Canula
26
Method
2,885,000
PAL
Manual Canula
1,593,333
2,380,000
890,000
910,000
Age
varian
Sex
: Female greater in number of cell
yield count than male
Harvesting site : ADSc from flank greater in
number of cell yield count than from abdomen
Canula type : cell yield from manual canula
lipoaspirate greater in number than PAL
lipoaspirate.
Characterization of ADSc
tissue regeneration
The secretome includes angiogenic factors
(VEGF), anti-apoptotic factors (IGF-1),
hematopoietic factors (colony stimulating
factors & interleukins), & HGF
ADSc application on
esthetics
Soft tissue augmentation
The augmentation of breast was successful &
Conclusion
in regenerative medicine
Reference
Ther 2003;3:705713
Honga, S. J; Traktuev, D. O; and Marcha, K.L. Therapeutic potential
of adipose-derived stem cells in vascular growth and tissue repair.
Current Opinion in Organ Transplantation 2010:15:8691
Jin Jin, H. et al. Comparative Analysis of Human Mesenchymal Stem
Cells from Bone Marrow, Adipose Tissue, and Umbilical Cord Blood
as Sources of Cell Therapy. Int. J. Mol. Sci. 2013:14:17986-18001
Jeffrey M. Gimble, Adam J. Katz and Bruce A. Bunnell. AdiposeDerived Stem Cells for Regenerative Medicine. Journal of American
Heart Association. 2007;100:1249-1260
Lindroos, B.; Suuronen, B & Miettinen, S. The Potential of Adipose
Stem Cells in Regenerative Medicine. Stem Cell Rev and Rep,
Springer Science media. 2010
Madonna, R., Geng, Y-G., and Caterina, R. Adipose Tissue-Derived
Stem Cells : Characterization and Potential for Cardiovascular
Repair. Journal of American Heart Association. 2009:29:1723-1729
Thankyou