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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 ASC isolation relative to duration of cryopreservation. (a) ASCs were isolated from 32 patients whose adipose tissue was cryopreserved for varying amounts of time (range 2–1159 days, average 596.4 × 104 ± 369.9 × 104 cells/g tissue). Live ASCs isolated ranged from 0 to 95.5 × 104 cells/g adipose, average 63.8 × 104 ± 30.6 × 104 cells/g tissue, showing a trend toward decreased live ASC isolation with increasing ASC cryopreservation duration. (b) Live cell count was compared relative to cryopreservation duration in 3 cohort groups: <1 year (N = 10), 1-2 years (N = 5), and >2 years (N = 17). A significant decrease in live ASC isolation was observed between the >2 years and <1 year cryopreservation duration cohorts; P = 0.0003. (c) Patient age was compared for each of the cryopreservation cohort groups and no significant differences were found between any group; P > 0.05.
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fig1: Live ASC isolation relative to duration of cryopreservation. (a) ASCs were isolated from 32 patients whose adipose tissue was cryopreserved for varying amounts of time (range 2–1159 days, average 596.4 × 104 ± 369.9 × 104 cells/g tissue). Live ASCs isolated ranged from 0 to 95.5 × 104 cells/g adipose, average 63.8 × 104 ± 30.6 × 104 cells/g tissue, showing a trend toward decreased live ASC isolation with increasing ASC cryopreservation duration. (b) Live cell count was compared relative to cryopreservation duration in 3 cohort groups: <1 year (N = 10), 1-2 years (N = 5), and >2 years (N = 17). A significant decrease in live ASC isolation was observed between the >2 years and <1 year cryopreservation duration cohorts; P = 0.0003. (c) Patient age was compared for each of the cryopreservation cohort groups and no significant differences were found between any group; P > 0.05.

Mentions: Live ASCs isolated ranged from 0 to 5.9 × 104 cells/g adipose tissue harvested, averaging 2.95 × 104 ± 2.5 × 104 cells/g tissue. ASC isolation from frozen tissue of cohorts <1 year, 1-2 years, and >2 years had average live cell count values of 4.06 × 104 ± 1.36 × 104 cells/g tissue, 2.29 × 104 ± 2.24 × 104 cells/g tissue, and 1.87 × 104 ± 1.12 × 104 cells/g tissue, respectively. We observed an inverse relationship between the number of live ASCs isolated in relation to the number of days the tissue was frozen (R2 = 0.1093, Figure 1(a)). There was a significantly greater number of live ASCs isolated from samples frozen <1 year compared to those frozen >2 years (P = 0.0003). No significant differences were found between other groups (Figure 1(b)). Multivariate regression analysis demonstrated no significant difference in the number of cells isolated relative to patient age or other demographic variables (P > 0.05). In contrast, cryopreservation length was independently associated with initial number of cells isolated irrespective of patient age (P < 0.001, Figure 1(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 ASC isolation relative to duration of cryopreservation. (a) ASCs were isolated from 32 patients whose adipose tissue was cryopreserved for varying amounts of time (range 2–1159 days, average 596.4 × 104 ± 369.9 × 104 cells/g tissue). Live ASCs isolated ranged from 0 to 95.5 × 104 cells/g adipose, average 63.8 × 104 ± 30.6 × 104 cells/g tissue, showing a trend toward decreased live ASC isolation with increasing ASC cryopreservation duration. (b) Live cell count was compared relative to cryopreservation duration in 3 cohort groups: <1 year (N = 10), 1-2 years (N = 5), and >2 years (N = 17). A significant decrease in live ASC isolation was observed between the >2 years and <1 year cryopreservation duration cohorts; P = 0.0003. (c) Patient age was compared for each of the cryopreservation cohort groups and no significant differences were found between any group; P > 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig1: Live ASC isolation relative to duration of cryopreservation. (a) ASCs were isolated from 32 patients whose adipose tissue was cryopreserved for varying amounts of time (range 2–1159 days, average 596.4 × 104 ± 369.9 × 104 cells/g tissue). Live ASCs isolated ranged from 0 to 95.5 × 104 cells/g adipose, average 63.8 × 104 ± 30.6 × 104 cells/g tissue, showing a trend toward decreased live ASC isolation with increasing ASC cryopreservation duration. (b) Live cell count was compared relative to cryopreservation duration in 3 cohort groups: <1 year (N = 10), 1-2 years (N = 5), and >2 years (N = 17). A significant decrease in live ASC isolation was observed between the >2 years and <1 year cryopreservation duration cohorts; P = 0.0003. (c) Patient age was compared for each of the cryopreservation cohort groups and no significant differences were found between any group; P > 0.05.
Mentions: Live ASCs isolated ranged from 0 to 5.9 × 104 cells/g adipose tissue harvested, averaging 2.95 × 104 ± 2.5 × 104 cells/g tissue. ASC isolation from frozen tissue of cohorts <1 year, 1-2 years, and >2 years had average live cell count values of 4.06 × 104 ± 1.36 × 104 cells/g tissue, 2.29 × 104 ± 2.24 × 104 cells/g tissue, and 1.87 × 104 ± 1.12 × 104 cells/g tissue, respectively. We observed an inverse relationship between the number of live ASCs isolated in relation to the number of days the tissue was frozen (R2 = 0.1093, Figure 1(a)). There was a significantly greater number of live ASCs isolated from samples frozen <1 year compared to those frozen >2 years (P = 0.0003). No significant differences were found between other groups (Figure 1(b)). Multivariate regression analysis demonstrated no significant difference in the number of cells isolated relative to patient age or other demographic variables (P > 0.05). In contrast, cryopreservation length was independently associated with initial number of cells isolated irrespective of patient age (P < 0.001, Figure 1(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.