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Successful isolation of viable adipose-derived stem cells from human adipose tissue subject to long-term cryopreservation: positive implications for adult stem cell-based therapeutics in patients of advanced age.

Devitt SM, Carter CM, Dierov R, Weiss S, Gersch RP, Percec I - Stem Cells Int (2015)

Bottom Line: Significantly more viable cells were initially isolated from tissue cryopreserved <1 year than from tissue cryopreserved >2 years, irrespective of patient age.Mesenchymal stem cell markers were maintained in all cohorts tested throughout the duration of the study.Patient age does not significantly impact stem cell isolation, viability, or growth.

View Article: PubMed Central - PubMed

Affiliation: Thomas Jefferson University Hospital, 132 S 10th Street No. 763J, Philadelphia, PA 19107, USA.

ABSTRACT
We examined cell isolation, viability, and growth in adipose-derived stem cells harvested from whole adipose tissue subject to different cryopreservation lengths (2-1159 days) from patients of varying ages (26-62 years). Subcutaneous abdominal adipose tissue was excised during abdominoplasties and was cryopreserved. The viability and number of adipose-derived stem cells isolated were measured after initial isolation and after 9, 18, and 28 days of growth. Data were analyzed with respect to cryopreservation duration and patient age. Significantly more viable cells were initially isolated from tissue cryopreserved <1 year than from tissue cryopreserved >2 years, irrespective of patient age. However, this difference did not persist with continued growth and there were no significant differences in cell viability or growth at subsequent time points with respect to cryopreservation duration or patient age. Mesenchymal stem cell markers were maintained in all cohorts tested throughout the duration of the study. Consequently, longer cryopreservation negatively impacts initial live adipose-derived stem cell isolation; however, this effect is neutralized with continued cell growth. Patient age does not significantly impact stem cell isolation, viability, or growth. Cryopreservation of adipose tissue is an effective long-term banking method for isolation of adipose-derived stem cells in patients of varying ages.

No MeSH data available.


Live ASCs during extended cell growth relative to patient age. ASCs from each patient were plated to a density of 1 × 105 cells/well and viability was assayed after (a) 9, (b) 18, and (c) 28 days to characterize the effect of patient age on continued ASC viability. We observed sustained ASC viability irrespective of patient age. Cell counts were compared relative to patients across 4 cohort groups: <40 years (N = 12) versus ≥40 years (N = 20); <50 years (N = 23) versus ≥50 years (N = 9); and <40 years (N = 12) versus ≥50 years (N = 9). Live cell number was not found to be significantly different in any of the comparisons.
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fig6: Live ASCs during extended cell growth relative to patient age. ASCs from each patient were plated to a density of 1 × 105 cells/well and viability was assayed after (a) 9, (b) 18, and (c) 28 days to characterize the effect of patient age on continued ASC viability. We observed sustained ASC viability irrespective of patient age. Cell counts were compared relative to patients across 4 cohort groups: <40 years (N = 12) versus ≥40 years (N = 20); <50 years (N = 23) versus ≥50 years (N = 9); and <40 years (N = 12) versus ≥50 years (N = 9). Live cell number was not found to be significantly different in any of the comparisons.

Mentions: ASCs from each patient were plated to a density of 1 × 105 cells/well and counted 9, 18, and 28 days later to examine and to characterize the effect of patient age on continued ASC growth. We observed no significant differences in ASC growth relative to patient age when the average cell number was compared for the following age groups: (1) <40 years versus ≥40 year cohorts after 9 days of growth (1.73 × 105 ± 0.7 × 105 cells and 2.14 × 105 ± 0.88 × 105 cells, resp.), 18 days of growth (3.85 × 105 ± 2.13 × 105 cells and 4.85 × 105 ± 2.59 × 105 cells, resp.), or 28 days of growth (9.09 × 105 ± 3.22 × 105 cells and 9.49 × 105 ± 5.2 × 105 cells, resp.) in culture; (2) <50 years versus ≥50 year cohorts after 9 days of growth (1.9 × 105 ± 0.75 × 105 cells and 2.29 × 105 ± 1.04 × 105 cells, resp.), 18 days of growth (4.44 × 105 ± 2.44 × 105 cells and 4.78 × 105 ± 2.63 × 105 cells, resp.), or 28 days of growth (9.92 × 105 ± 4.49 × 105 cells and 6.41 × 105 ± 2.05 × 105 cells, resp.) in culture; (3) <40 years versus ≥50 cohorts after 9 days of growth (1.73 × 105 ± 0.7 × 105 cells and 2.29 × 105 ± 1.04 × 105 cells, resp.), 18 days of growth (3.85 × 105 ± 2.13 × 105 cells and 4.78 × 105 ± 2.63 × 105 cells, resp.), or 28 days of growth (9.09 × 105 ± 3.22 × 105 cells and 6.41 × 105 ± 2.05 × 105 cells, resp.; Figures 6(a)–6(c)).


Successful isolation of viable adipose-derived stem cells from human adipose tissue subject to long-term cryopreservation: positive implications for adult stem cell-based therapeutics in patients of advanced age.

Devitt SM, Carter CM, Dierov R, Weiss S, Gersch RP, Percec I - Stem Cells Int (2015)

Live ASCs during extended cell growth relative to patient age. ASCs from each patient were plated to a density of 1 × 105 cells/well and viability was assayed after (a) 9, (b) 18, and (c) 28 days to characterize the effect of patient age on continued ASC viability. We observed sustained ASC viability irrespective of patient age. Cell counts were compared relative to patients across 4 cohort groups: <40 years (N = 12) versus ≥40 years (N = 20); <50 years (N = 23) versus ≥50 years (N = 9); and <40 years (N = 12) versus ≥50 years (N = 9). Live cell number was not found to be significantly different in any of the comparisons.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4402176&req=5

fig6: Live ASCs during extended cell growth relative to patient age. ASCs from each patient were plated to a density of 1 × 105 cells/well and viability was assayed after (a) 9, (b) 18, and (c) 28 days to characterize the effect of patient age on continued ASC viability. We observed sustained ASC viability irrespective of patient age. Cell counts were compared relative to patients across 4 cohort groups: <40 years (N = 12) versus ≥40 years (N = 20); <50 years (N = 23) versus ≥50 years (N = 9); and <40 years (N = 12) versus ≥50 years (N = 9). Live cell number was not found to be significantly different in any of the comparisons.
Mentions: ASCs from each patient were plated to a density of 1 × 105 cells/well and counted 9, 18, and 28 days later to examine and to characterize the effect of patient age on continued ASC growth. We observed no significant differences in ASC growth relative to patient age when the average cell number was compared for the following age groups: (1) <40 years versus ≥40 year cohorts after 9 days of growth (1.73 × 105 ± 0.7 × 105 cells and 2.14 × 105 ± 0.88 × 105 cells, resp.), 18 days of growth (3.85 × 105 ± 2.13 × 105 cells and 4.85 × 105 ± 2.59 × 105 cells, resp.), or 28 days of growth (9.09 × 105 ± 3.22 × 105 cells and 9.49 × 105 ± 5.2 × 105 cells, resp.) in culture; (2) <50 years versus ≥50 year cohorts after 9 days of growth (1.9 × 105 ± 0.75 × 105 cells and 2.29 × 105 ± 1.04 × 105 cells, resp.), 18 days of growth (4.44 × 105 ± 2.44 × 105 cells and 4.78 × 105 ± 2.63 × 105 cells, resp.), or 28 days of growth (9.92 × 105 ± 4.49 × 105 cells and 6.41 × 105 ± 2.05 × 105 cells, resp.) in culture; (3) <40 years versus ≥50 cohorts after 9 days of growth (1.73 × 105 ± 0.7 × 105 cells and 2.29 × 105 ± 1.04 × 105 cells, resp.), 18 days of growth (3.85 × 105 ± 2.13 × 105 cells and 4.78 × 105 ± 2.63 × 105 cells, resp.), or 28 days of growth (9.09 × 105 ± 3.22 × 105 cells and 6.41 × 105 ± 2.05 × 105 cells, resp.; Figures 6(a)–6(c)).

Bottom Line: Significantly more viable cells were initially isolated from tissue cryopreserved <1 year than from tissue cryopreserved >2 years, irrespective of patient age.Mesenchymal stem cell markers were maintained in all cohorts tested throughout the duration of the study.Patient age does not significantly impact stem cell isolation, viability, or growth.

View Article: PubMed Central - PubMed

Affiliation: Thomas Jefferson University Hospital, 132 S 10th Street No. 763J, Philadelphia, PA 19107, USA.

ABSTRACT
We examined cell isolation, viability, and growth in adipose-derived stem cells harvested from whole adipose tissue subject to different cryopreservation lengths (2-1159 days) from patients of varying ages (26-62 years). Subcutaneous abdominal adipose tissue was excised during abdominoplasties and was cryopreserved. The viability and number of adipose-derived stem cells isolated were measured after initial isolation and after 9, 18, and 28 days of growth. Data were analyzed with respect to cryopreservation duration and patient age. Significantly more viable cells were initially isolated from tissue cryopreserved <1 year than from tissue cryopreserved >2 years, irrespective of patient age. However, this difference did not persist with continued growth and there were no significant differences in cell viability or growth at subsequent time points with respect to cryopreservation duration or patient age. Mesenchymal stem cell markers were maintained in all cohorts tested throughout the duration of the study. Consequently, longer cryopreservation negatively impacts initial live adipose-derived stem cell isolation; however, this effect is neutralized with continued cell growth. Patient age does not significantly impact stem cell isolation, viability, or growth. Cryopreservation of adipose tissue is an effective long-term banking method for isolation of adipose-derived stem cells in patients of varying ages.

No MeSH data available.