Cryopreservation of whole adipose tissue for future use in regenerative medicine

Mahmood S. Choudhery, Michael Badowski, Angela Muise, John Pierce, David T. Harris

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Background Human adipose tissue (AT) is an ideal stem cell source for autologous cell-based therapies. The preferred setting for tissue engineering and regenerative medicine applications is the availability of clinically acceptable off-the-shelf cells and cell products. As AT is not always available for use, cryopreserved tissue represents an alternative approach. The aim of the present study was to compare the different properties of mesenchymal stem cells (MSCs) isolated from cryopreserved AT. We have measured cell recovery, viability, phenotype, proliferative potential, and differentiation into mesenchymal (adipogenic, osteogenic, chondrogenic) and nonmesenchymal (neuron-like cells) lineages. Materials and methods AT (n = 10) was harvested from donors and either processed fresh or cryopreserved in liquid nitrogen dewars. Both fresh and thawed tissues were enzymatically digested. MSCs were analyzed by fluorescence-activated cell sorting for CD3, CD14, CD19, CD34, CD44, CD45, CD73, CD90, and CD105 expression. Growth characteristics of both groups were investigated for population doublings, doubling time, saturation density, and plating efficiency. MSCs derived from fresh and thawed tissues were assessed for differentiation potential both qualitatively and quantitatively. Results Adherent cells from fresh and thawed tissues displayed similar fibroblastic morphology. Cryopreservation did not alter expression of phenotypic markers. Similarly, the proliferative potential of MSCs was not compromised by cryopreservation. Furthermore, cryopreservation did not alter the differentiation capability of MSCs as determined with histochemistry, immunofluorescence, and real time reverse transcriptase-polymerase chain reaction. Conclusions We conclude that human AT could be successfully cryopreserved for future clinical application and the recovered MSCs were equivalent in functionality to the freshly processed MSCs.

Original languageEnglish (US)
Pages (from-to)24-35
Number of pages12
JournalJournal of Surgical Research
Volume187
Issue number1
DOIs
StatePublished - Mar 2014

Fingerprint

Regenerative Medicine
Cryopreservation
Mesenchymal Stromal Cells
Adipose Tissue
Cell Lineage
Tissue Engineering
Cell- and Tissue-Based Therapy
Reverse Transcriptase Polymerase Chain Reaction
Fluorescent Antibody Technique
Real-Time Polymerase Chain Reaction
Cell Survival
Flow Cytometry
Nitrogen
Stem Cells
Phenotype
Neurons
Growth
Population

Keywords

  • Adipose tissue
  • Cryopreservation
  • MSC
  • Regenerative medicine
  • Tissue engineering

ASJC Scopus subject areas

  • Surgery

Cite this

Cryopreservation of whole adipose tissue for future use in regenerative medicine. / Choudhery, Mahmood S.; Badowski, Michael; Muise, Angela; Pierce, John; Harris, David T.

In: Journal of Surgical Research, Vol. 187, No. 1, 03.2014, p. 24-35.

Research output: Contribution to journalArticle

Choudhery, Mahmood S. ; Badowski, Michael ; Muise, Angela ; Pierce, John ; Harris, David T. / Cryopreservation of whole adipose tissue for future use in regenerative medicine. In: Journal of Surgical Research. 2014 ; Vol. 187, No. 1. pp. 24-35.
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abstract = "Background Human adipose tissue (AT) is an ideal stem cell source for autologous cell-based therapies. The preferred setting for tissue engineering and regenerative medicine applications is the availability of clinically acceptable off-the-shelf cells and cell products. As AT is not always available for use, cryopreserved tissue represents an alternative approach. The aim of the present study was to compare the different properties of mesenchymal stem cells (MSCs) isolated from cryopreserved AT. We have measured cell recovery, viability, phenotype, proliferative potential, and differentiation into mesenchymal (adipogenic, osteogenic, chondrogenic) and nonmesenchymal (neuron-like cells) lineages. Materials and methods AT (n = 10) was harvested from donors and either processed fresh or cryopreserved in liquid nitrogen dewars. Both fresh and thawed tissues were enzymatically digested. MSCs were analyzed by fluorescence-activated cell sorting for CD3, CD14, CD19, CD34, CD44, CD45, CD73, CD90, and CD105 expression. Growth characteristics of both groups were investigated for population doublings, doubling time, saturation density, and plating efficiency. MSCs derived from fresh and thawed tissues were assessed for differentiation potential both qualitatively and quantitatively. Results Adherent cells from fresh and thawed tissues displayed similar fibroblastic morphology. Cryopreservation did not alter expression of phenotypic markers. Similarly, the proliferative potential of MSCs was not compromised by cryopreservation. Furthermore, cryopreservation did not alter the differentiation capability of MSCs as determined with histochemistry, immunofluorescence, and real time reverse transcriptase-polymerase chain reaction. Conclusions We conclude that human AT could be successfully cryopreserved for future clinical application and the recovered MSCs were equivalent in functionality to the freshly processed MSCs.",
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AB - Background Human adipose tissue (AT) is an ideal stem cell source for autologous cell-based therapies. The preferred setting for tissue engineering and regenerative medicine applications is the availability of clinically acceptable off-the-shelf cells and cell products. As AT is not always available for use, cryopreserved tissue represents an alternative approach. The aim of the present study was to compare the different properties of mesenchymal stem cells (MSCs) isolated from cryopreserved AT. We have measured cell recovery, viability, phenotype, proliferative potential, and differentiation into mesenchymal (adipogenic, osteogenic, chondrogenic) and nonmesenchymal (neuron-like cells) lineages. Materials and methods AT (n = 10) was harvested from donors and either processed fresh or cryopreserved in liquid nitrogen dewars. Both fresh and thawed tissues were enzymatically digested. MSCs were analyzed by fluorescence-activated cell sorting for CD3, CD14, CD19, CD34, CD44, CD45, CD73, CD90, and CD105 expression. Growth characteristics of both groups were investigated for population doublings, doubling time, saturation density, and plating efficiency. MSCs derived from fresh and thawed tissues were assessed for differentiation potential both qualitatively and quantitatively. Results Adherent cells from fresh and thawed tissues displayed similar fibroblastic morphology. Cryopreservation did not alter expression of phenotypic markers. Similarly, the proliferative potential of MSCs was not compromised by cryopreservation. Furthermore, cryopreservation did not alter the differentiation capability of MSCs as determined with histochemistry, immunofluorescence, and real time reverse transcriptase-polymerase chain reaction. Conclusions We conclude that human AT could be successfully cryopreserved for future clinical application and the recovered MSCs were equivalent in functionality to the freshly processed MSCs.

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