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Scalable efficient expansion of mesenchymal stem cells in xeno free media using commercially available reagents.

Riordan NH, Madrigal M, Reneau J, de Cupeiro K, Jiménez N, Ruiz S, Sanchez N, Ichim TE, Silva F, Patel AN - J Transl Med (2015)

Bottom Line: The rapid clinical translation of mesenchymal stem cells (MSC) has resulted in the development of cell-based strategies for multiple indications.This study demonstrated equivalent or superior effects of human platelet lysate compared to standard FCS supplemented media, based on doubling rate, without loss of identity or function, as demonstrated with flow cytometry characterization.Differentiation into osteocytes, adipocytes and chondrocytes was comparable from cells expanded in either media supplement.

View Article: PubMed Central - PubMed

Affiliation: Medistem Panama, Inc., Building 221, City of Knowledge, Clayton, Panama City, Republic of Panama. nhriordan@gmail.com.

ABSTRACT

Background: The rapid clinical translation of mesenchymal stem cells (MSC) has resulted in the development of cell-based strategies for multiple indications. Unfortunately one major barrier to widespread implementation of MSC-based therapies is the limited supply of fetal calf serum (FCS) used to expand cells to therapeutic numbers. Additionally, the xenogeneic element of fetal calf serum has been previously demonstrated to stimulate antibody mediated reactions and in some cases sensitization leading to anaphylaxis.

Method: XcytePLUS™ media, a human platelet lysate based product, was used to supplement the culture medium at 5, 7.5 and 10% and compared to fetal calf serum at 10%, for human umbilical cord MSC expansion. Properties of the expanded cells were investigated.

Results: This study demonstrated equivalent or superior effects of human platelet lysate compared to standard FCS supplemented media, based on doubling rate, without loss of identity or function, as demonstrated with flow cytometry characterization. Differentiation into osteocytes, adipocytes and chondrocytes was comparable from cells expanded in either media supplement.

Conclusions: These data support the implementation of human platelet lysate supplemented media as an alternative to xenogeneic containing preparations which may lead to safer MSC products with therapeutic uses.

No MeSH data available.


Related in: MedlinePlus

Morphology of WJ-MSC is not altered by supplementation of culture media with XcytePLUS™ compared to Fetal Calf Serum (FCS). 10X inverted phase contrast microscope images of WJ-MSC cultured in a 10% FCS, b 5% XcytePLUS™, c 7.5% XcytePLUS™ and d 10% XcytePLUS™ on passage 5.
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Fig1: Morphology of WJ-MSC is not altered by supplementation of culture media with XcytePLUS™ compared to Fetal Calf Serum (FCS). 10X inverted phase contrast microscope images of WJ-MSC cultured in a 10% FCS, b 5% XcytePLUS™, c 7.5% XcytePLUS™ and d 10% XcytePLUS™ on passage 5.

Mentions: WJ-MSC after 5 passages were plated in media containing 5, 7.5 and 10% XcytePLUS™, or 10% FCS supplemented media and cultured for 9 days. Morphological differences were not found between the treatment groups (Figure 1). Significantly higher number of WJ-MSC were collected on day 9 with the 10% XcytePLUS™ supplemented media (average 1.4 × 107) as compared to 10% FCS media (average 7.5 × 106) (p < 0.05). Average numbers are represented in Figure 2. Furthermore, population doublings in 9 days were 8.37 for cells supplemented with 10% XcytePLUS™ and 7.29 using 10% FCS. Doublings per day did not show statistical difference when different lots of XcytePLUS™ were used (Figure 3). Furthermore, doubling rate experiments performed on the same four lots in different laboratories did not show substantial differences (Figure 4).Figure 1


Scalable efficient expansion of mesenchymal stem cells in xeno free media using commercially available reagents.

Riordan NH, Madrigal M, Reneau J, de Cupeiro K, Jiménez N, Ruiz S, Sanchez N, Ichim TE, Silva F, Patel AN - J Transl Med (2015)

Morphology of WJ-MSC is not altered by supplementation of culture media with XcytePLUS™ compared to Fetal Calf Serum (FCS). 10X inverted phase contrast microscope images of WJ-MSC cultured in a 10% FCS, b 5% XcytePLUS™, c 7.5% XcytePLUS™ and d 10% XcytePLUS™ on passage 5.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4504159&req=5

Fig1: Morphology of WJ-MSC is not altered by supplementation of culture media with XcytePLUS™ compared to Fetal Calf Serum (FCS). 10X inverted phase contrast microscope images of WJ-MSC cultured in a 10% FCS, b 5% XcytePLUS™, c 7.5% XcytePLUS™ and d 10% XcytePLUS™ on passage 5.
Mentions: WJ-MSC after 5 passages were plated in media containing 5, 7.5 and 10% XcytePLUS™, or 10% FCS supplemented media and cultured for 9 days. Morphological differences were not found between the treatment groups (Figure 1). Significantly higher number of WJ-MSC were collected on day 9 with the 10% XcytePLUS™ supplemented media (average 1.4 × 107) as compared to 10% FCS media (average 7.5 × 106) (p < 0.05). Average numbers are represented in Figure 2. Furthermore, population doublings in 9 days were 8.37 for cells supplemented with 10% XcytePLUS™ and 7.29 using 10% FCS. Doublings per day did not show statistical difference when different lots of XcytePLUS™ were used (Figure 3). Furthermore, doubling rate experiments performed on the same four lots in different laboratories did not show substantial differences (Figure 4).Figure 1

Bottom Line: The rapid clinical translation of mesenchymal stem cells (MSC) has resulted in the development of cell-based strategies for multiple indications.This study demonstrated equivalent or superior effects of human platelet lysate compared to standard FCS supplemented media, based on doubling rate, without loss of identity or function, as demonstrated with flow cytometry characterization.Differentiation into osteocytes, adipocytes and chondrocytes was comparable from cells expanded in either media supplement.

View Article: PubMed Central - PubMed

Affiliation: Medistem Panama, Inc., Building 221, City of Knowledge, Clayton, Panama City, Republic of Panama. nhriordan@gmail.com.

ABSTRACT

Background: The rapid clinical translation of mesenchymal stem cells (MSC) has resulted in the development of cell-based strategies for multiple indications. Unfortunately one major barrier to widespread implementation of MSC-based therapies is the limited supply of fetal calf serum (FCS) used to expand cells to therapeutic numbers. Additionally, the xenogeneic element of fetal calf serum has been previously demonstrated to stimulate antibody mediated reactions and in some cases sensitization leading to anaphylaxis.

Method: XcytePLUS™ media, a human platelet lysate based product, was used to supplement the culture medium at 5, 7.5 and 10% and compared to fetal calf serum at 10%, for human umbilical cord MSC expansion. Properties of the expanded cells were investigated.

Results: This study demonstrated equivalent or superior effects of human platelet lysate compared to standard FCS supplemented media, based on doubling rate, without loss of identity or function, as demonstrated with flow cytometry characterization. Differentiation into osteocytes, adipocytes and chondrocytes was comparable from cells expanded in either media supplement.

Conclusions: These data support the implementation of human platelet lysate supplemented media as an alternative to xenogeneic containing preparations which may lead to safer MSC products with therapeutic uses.

No MeSH data available.


Related in: MedlinePlus